From 2cd89cceb134319489541a4c7f2c3a47291ed80e Mon Sep 17 00:00:00 2001
From: Kathleen Susat <kathleen.susat@uni-bielefeld.de>
Date: Sat, 23 Nov 2024 11:04:49 +0100
Subject: [PATCH] figure description
---
src/contents/results.tsx | 30 +++++++++++++++---------------
1 file changed, 15 insertions(+), 15 deletions(-)
diff --git a/src/contents/results.tsx b/src/contents/results.tsx
index 796fd52a..ff6ed270 100644
--- a/src/contents/results.tsx
+++ b/src/contents/results.tsx
@@ -38,7 +38,7 @@ export function Results() {
<H5 text="Conclusion"/>
<p>The microscopy data validates our proof of concept. Compared to our internal positive control, pDAS12124-preedited (see Figure 1), less cells co-transfected with pDAS12489 and pCMV-PE2 (see Figure 1) showed fluorescence. Contrary to our expectations, the technical transfection control with pZMB938 showed lower transfection efficiency. All negative controls showed no fluorescence.</p>
<ThreeVertical
- description="Microscopy of HEK293 72h post transfection with lipofectamine 2000. Transfection with technical positive control pZMB938, internal positive control pDAS12124-preedited, co-transfection of pDAS12489 with pCMV-PE2, NTC, PE2 as control and pDAS12489 as control. All controls are negative and both positve controls as well as pDAS12489+pCMV-PE2 show fluorescence signals."
+ description="Microscopy of HEK293 72h post transfection with lipofectamine 2000. Transfection with technical positive control pZMB938, internal positive control pDAS12124-preedited, co-transfection of pDAS12489 with pCMV-PE2, NTC, PE2 as control and pDAS12489 as control. All controls are negative and both positve controls as well as pDAS12489+pCMV-PE2 show fluorescence signals. BF = Brightfield"
num={1}
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/bild1-1.png"
alt1="pZMB"
@@ -52,7 +52,7 @@ export function Results() {
<H5 text="Conclusion"/>
<p>All 4 different transfection conditions were done with pZMB938 and showed good results, but best result were done when lipofectamine 2000 was diluted 1:10 and 1000 ng DNA was transfected.</p>
<TwoVertical
- description="Microscopy of HEK293 72h post transfection with lipofectamin 2000. Transfection with 1:10 or 1:25 diluted lipofectamine and 800 ng or 1000 ng of out technical positive control pZMB938."
+ description="Microscopy of HEK293 72h post transfection with lipofectamin 2000. Transfection with 1:10 or 1:25 diluted lipofectamine and 800 ng or 1000 ng of out technical positive control pZMB938. BF = Brightfield"
num={2}
bg="white"
alt1="1000ng DNA with 1:10 and 1:25 Lipofectamine 2000"
@@ -64,7 +64,7 @@ export function Results() {
<H5 text="Conclusion"/>
<p>Internal control and technical control showed higher transfection efficiency then in previous experiments, therefore transfection with lipofectamine 3000 seems to be more efficient than transfection with lipofectamine 2000. The fluorescence of pDAS12189+pCMV-PE2 was still quite low. All negative controls are showed no fluorescence.</p>
<ThreeVertical
- description="Microscopy of HEK293 72h post transfection with lipofectamine 2000. Transfection with technical positive control pZMB938, internal positive control pDAS12124-preedited, co-transfection of pDAS12489 with pCMV-PE2, NTC, PE2 as control and pDAS12489 as control. All controls are negative and both positve controls as well as pDAS12489+pCMV-PE2 show fluorescence signals."
+ description="Microscopy of HEK293 72h post transfection with lipofectamine 2000. Transfection with technical positive control pZMB938, internal positive control pDAS12124-preedited, co-transfection of pDAS12489 with pCMV-PE2, NTC, PE2 as control and pDAS12489 as control. All controls are negative and both positve controls as well as pDAS12489+pCMV-PE2 show fluorescence signals. BF = Brightfield"
num={3}
bg="white"
alt1="pZMB and pDAS12124"
@@ -77,7 +77,7 @@ export function Results() {
<H5 text="Conclusion"/>
<p>The results of the test were not as good as expected. Nearly no transfection efficiency was visible. This could be due to too old HEK293 cells</p>
<TwoVertical
- description="Microscopy of HEK293 72h post transfection with lipofectaine 3000. Transfection of 500 ng or 1000 ng of our technical positive control pZMB938 with 1 µl or 1.5 µl of lipofectamine 3000."
+ description="Microscopy of HEK293 72h post transfection with lipofectaine 3000. Transfection of 500 ng or 1000 ng of our technical positive control pZMB938 with 1 µl or 1.5 µl of lipofectamine 3000. BF = Brightfield"
num={4}
bg="white"
alt1="500ng DNA 1 µl or 1.5 µl of lipofectamine 3000"
@@ -93,11 +93,11 @@ export function Results() {
num={5}
bg="white"
alt1="Microscopy of HEK293 72h post transfection with lipofectamine 3000 with 1000 ng or 1500 ng technical positive control pZMB938. Both transfections show fluorescence signals"
- description="Microscopy of HEK293 72h post transfection with lipofectamine 3000 with 1000 ng or 1500 ng technical positive control pZMB938. Both transfections show fluorescence signals"
+ description="Microscopy of HEK293 72h post transfection with lipofectamine 3000 with 1000 ng or 1500 ng technical positive control pZMB938. Both transfections show fluorescence signals. BF = Brightfield"
/>
<TwoVertical
- description="Microscopy of HEK293 72h post transfection with CaCl2 with 500 ng, 1000 ng or 1500 ng pZMB938. All transfections show fluorescence signals."
+ description="Microscopy of HEK293 72h post transfection with CaCl2 with 500 ng, 1000 ng or 1500 ng pZMB938. All transfections show fluorescence signals. BF = Brightfield"
num={6}
bg="white"
alt1=""
@@ -110,7 +110,7 @@ export function Results() {
<p>The transfection efficiency was much better. Our proof-of-concept was working correctly. The reporter system pDAS12189 only led to production of a fluorescent signal when co transfected with a prime editing complex as pCMV-PE2.</p>
<TwoVertical
alt1=""
- description="Microscopy of HEK293 72h post transfection with lipofectamine 2000. Transfection with technical positive control pZMB938, internal positive control pDAS12124-preedited, co-transfection of pDAS12489 with pCMV-PE2, NTC, PE2 as control and pDAS12489 as control. All controls are negative and both positve controls as well as pDAS12489+pCMV-PE2 show fluorescence signals."
+ description="Microscopy of HEK293 72h post transfection with lipofectamine 2000. Transfection with technical positive control pZMB938, internal positive control pDAS12124-preedited, co-transfection of pDAS12489 with pCMV-PE2, NTC, PE2 as control and pDAS12489 as control. All controls are negative and both positve controls as well as pDAS12489+pCMV-PE2 show fluorescence signals. BF = Brightfield"
num={7}
bg="white"
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/bild7-1.png"
@@ -124,7 +124,7 @@ export function Results() {
<H5 text="Conclusion"/>
<p>The Flow Cytometry analysis shows that pegRNA without silent edits (pegRNA1) had a 2.05 times higher transfection efficiency than pegRNA with silent edits (pegRNA2).</p>
<TwoHorizontal
- description="Flow Cytometry analysis of pegRNAs with and without silent edits."
+ description="Flow cytometry analysis of pegRNAs with and without silent edits. Histograms of cell count applied against fluorescence intensity of healthy HEK293 cells (left) with untransfected cells as negative control, pDAS12124 pre-edited as internal positive control and pegRNAs with (pegRNA1) and without (pegRNA2) silent edits 72 h after transfection. The portion of fluorescent cells is normalized to the internal positive control (right)."
num={8}
bg="white"
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/se-nose.png"
@@ -143,7 +143,7 @@ export function Results() {
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/bild9.png"
num={9}
alt1="Flow Cytometry results of all screened pegRNAs"
- description="Flow Cytometry results of all screened pegRNAs"
+ description="Percentage of fluorescent HEK293 cells 72 h after transfection with various pegRNAs (pegRNA1-14) normalized to pDAS12124 pre-edited as internal positive control as result of flow cytometry analysis."
/>
</div>
</div>
@@ -154,7 +154,7 @@ export function Results() {
<p>Transfection of CFBE41o- with pDAS12124-preedited was successful (see Figure 10). After 24 hours a successful co transfection of pPEAR_CFTR with PE6c and pegRNA4 was visible, although the transfection efficiency was really bad (see Figure 10).</p>
<TwoVertical
alt1=""
- description="Microscopy results after 24h or 48h. Transfection of pDAS12124-preedited with lipofectamine 3000 was successfully done in CFBE41o- cell line and visible after 48h. CFBE41o- cell line was transfected with pDAS-IDT with Lipofectamine 3000 and afterwards with LNPs including PE6c and pegRNA4 and was after 24h fluorescence visible."
+ description="Microscopy results after 24h or 48h. Transfection of pDAS12124-preedited with lipofectamine 3000 was successfully done in CFBE41o- cell line and visible after 48h. CFBE41o- cell line was transfected with pDAS-IDT with Lipofectamine 3000 and afterwards with LNPs including PE6c and pegRNA4 and was after 24h fluorescence visible. BF = Brightfield"
num={10}
bg="white"
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/bild10-1.png"
@@ -163,7 +163,7 @@ export function Results() {
<p>Moreover transfection was conducted in human nasal epithelial cells (hNECs) in Air-liquid interface cultures as well as apical-out airway organoids (see Figure 11). No fluorescence was visible. </p>
<TwoVertical
alt1=""
- description="Microscopy of HEK 72h post transfection with lipofectamine 3000. Co-transfection of pPEAR_CFTR with PE6c and pegRNA4. Both show no fluorescence signals."
+ description="Microscopy of HEK 72h post transfection with lipofectamine 3000. Co-transfection of pPEAR_CFTR with PE6c and pegRNA4. Both show no fluorescence signals. BF = Brightfield"
num={11}
bg="white"
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/ali-tr.png"
@@ -178,7 +178,7 @@ export function Results() {
<div className="col">
<TwoVertical
alt1=""
- description="Microscopy of HEK 72h post transfection with lipofectamine 3000. Co-transfection of pDAS12489 with pCMV-PE2 or pDAS12489 with LV-PE_CO-Mini. Both show fluorescence signals."
+ description="Microscopy of HEK 72h post transfection with lipofectamine 3000. Co-transfection of pDAS12489 with pCMV-PE2 or pDAS12489 with LV-PE_CO-Mini. Both show fluorescence signals. BF = Brightfield"
num={12}
bg="white"
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/bild11-1.png"
@@ -192,7 +192,7 @@ export function Results() {
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/pe2-pe-co.png"
num={13}
bg="white"
- description="Flow Cytometry analysis to compare prime editing complexes PE2 and PE_CO-Mini"
+ description="Flow Cytometry analysis to compare prime editing complexes PE2 and PE_CO-Mini. Histograms of cell count applied against fluorescence intensity of healthy HEK293 cells (left) with untransfected cells as negative control, pDAS12124 pre-edited as internal positive control 72 h after transfection."
/>
</div>
</div>
@@ -206,13 +206,13 @@ export function Results() {
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/bild13.png"
num={14}
bg="white"
- description="Microscopy of HEK293 72h post transfection with lipofectamine 3000 and co transfection with pCMV-PE6c and pDAS12489."
+ description="Microscopy of HEK293 72h post transfection with lipofectamine 3000 and co transfection with pCMV-PE6c and pDAS12489. BF = Brightfield"
alt1="Microscopy of HEK293 72h post transfection with lipofectamine 3000 and co transfection with pCMV-PE6c and pDAS12489."
/>
<TwoHorizontal
alt1=""
- description="Flow Cytometry results for evaluation of PE6c performance."
+ description="Flow cytometry analysis for evaluation of performance of prime editor variants. Histograms of cell count applied against fluorescence intensity of healthy HEK293 cells (left) with untransfected cells as negative control, pDAS12124 pre-edited as internal positive control and PE6c, PE2 and PE2_Co-Mini 72 h after transfection. The portion of fluorescent cells is normalized to the internal positive control (right)."
num={15}
bg="white"
pic1="https://static.igem.wiki/teams/5247/photos/facs-results-mechanism/pe2-pe-co-pe6c.png"
--
GitLab