ELAV-Gal4;Tub-Gal80: Temperature switch at embryo stage
We phenocopied the results of the ELAV-Gal4 cross.
Now we are waiting for the results of the Temperature switch at the second instar larvae stage
![](https://lab.brembs.net/wp/wp-content/uploads/2020/12/first-experiment-1024x1007.jpg)
August
- Halfway in analyzing the Buridan data altogether (not in two batches)
- Maxiprepped plasmid for making FoxP protein at the klinikum
- Writing the thesis a little bit
- bought Fas-II antibody
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- data 1): FoxP-iB Heterozygous/Homozygous comparison with Stinger-GFP. This time i was cautious with everything: fly all the same age and sex, same larvae density, same number of copies of GFP
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/Picture1-1024x559.png)
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/Picture2-1024x571.png)
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/Picture3-1024x254.png)
I can not detect any difference between homozygous and heterozygous mutants. I also counted cells in IMARIS and I detected no difference in number or distribution.
- data 2): FoxP-iB Heterozygous/Homozygous comparison with CD8-GFP. This time i was cautious with everything: fly all the same age and sex, same larvae density, same number of copies of GFP
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/Picture4-1024x518.png)
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/Picture5-1024x547.png)
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/Picture6-1024x679.png)
I can see some differences but i do not know how to quantify/explain them. Also I am not sure if it is a problem of dissection/mounting. I am not convinced.
- data 3): nc82 staining on homo/hetero FoxP-iB. I do not see differences
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/Picture0.png)
Problem: I haven’t managed yet to make the FasII antibody work:
I have used a 1:10 concentration of primary antibody for one night and a 1:100 concentration of secondary antibody for 4 hours. I will ask someone
In the picture I have increased the gain a lot just to be sure. I could not see anything
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/MAX_Project.lif-Series005-1024x1024.jpg)
Tdc2-Gal4 + FoxP-LexA in VNC
![](https://lab.brembs.net/wp/wp-content/uploads/2020/06/Picture1-3-1024x510.png)
Green: Tdc2
No colocalization also in the VNC
Last week
- Crossed the flies for the course
- Almost finished the Buridan for C380
- Worked on the manuscript and supplemetary figures
- Done the FoxP reaction on induced embryo again because I was afraid after the different results Andi had last time
![](https://lab.brembs.net/wp/wp-content/uploads/2020/06/MAX_FoxP-Tubgal80ElavGal4-1024x1024.jpg)
qPCR-Joel
![](https://lab.brembs.net/wp/wp-content/uploads/2020/06/Untitled-1-1024x366.png)
Last week
- Made crosses for pupae TS
- Made cross for Tdc2-Gal4 and FoxP-LexA to see the VNC (I already had last week the pictures of the CB)
- Made crosses for ok107
- Made crosses for C380
After all those crosses will hatch I will have finished with the Buridan for now.