The first 3 images show the protocerebral bridge in FoxP mutated flies. In the second line the first picture shows the protocerebral bridge in a FoxP isoB-Gal4 fly. The last image is taken and modifies from the Virtual Fly Brain

Larvae ELAV (20x and 40x)

Adults (20x and 40x)

Larvae REPO (20x and 40x)

UNIVERSITY PART

First try for GABA antibody on FoxPisoB-Gal4 adult brain. The reaction is not optimal, there is a lot of background and the signal is too strong. We will have to try again changing conditions in order to be sure of any colocalization.

We also have found some FoxP-all_Isoforms-Gal4 homozygous, we thus wanted to see if there was any macro problem in the overall structure of the brain, but the structure looks fairly normal

UNIVERSITY PART

Larvae FoxPisoB-Gal4 + Bruch Pilot (20x and 40x)

KLINIKUM PART

I did the WB again in order to have a better and clearer image, additionally I made a coomassie staining and a silver staining in order to see if there is any contamination in the purified FoxP-IsoB protein

I also made a WB on Drosophila brains using the mouse serum. The signal is not clean but we can see a band of the appropriate size (60 kDa)

FoxP3955 flies were raised and compared to normal WTB flies. Reportedly, the Foxp mutants have a reduced flight performance as their total flight duration is decreased. This was also something I experienced. The problem seemed to be greater due to the heat in the flight simulator room, initial temperature was 27°C but increased to close to 30°C. I had troubles getting a large sample size enough (same number of Foxp and wtb were loaded into the flight simulator), heat-shock proteins and other stress-related behavior might be an issue. The genotype of the flies were known during the hooking of the flies but was later on concelead and flies were randomly distributed.  

cd8 GFP + Bruchpilot

Bruchpilot

Stinger GFP + REPO + ELAV

KLINIKUM PART

The serum from the forth injection recognize the FoxP recombinant protein. The band has the appropriate size

UNIVERSITY PART

I have exchanged the Hom1 domain that had a frameshift. Ready to inject again

KLINIKUM PART

The sera of the mice (expecially the one immunized with URE-10 peptide) show titer against FoxP recombinant protein. In september we can proceed with the hybridoma using the mouse immunized with ure-10

UNIVERSITY PART

Immunostaining against ELAV and BruchPilot in FoxP-IsoB-Gal4 flies (ELAV: 20x and 40x, BruchPilot 20x)

BruchPilot alone

Cell counting of 2 adult brains  for the Stinger staining in FoxP-IsoB-Gal4 flies. The expression is symmetrical and similar in the 2 brains. I am planning to do some more staining and counting at least 10 brains that I will compare with the FoxP all isoforms-Gal4