Extended Data Fig. 7: TH+ cell number in the PAG region remains unchanged between mutants and controls.
a) The representative image from a control mouse shows the PAG in the midbrain region around Bregma −2.488 (based on the Allen Brain Atlas). The image is immunostained for TH and visualised with a 647 nm far-red secondary antibody. Additionally, EBF3 in green (488 nm) is shown together with TH in the inset (b). Yellow arrowheads indicate a TH/EBF3 positive colocalised cell and white arrowheads indicate a non-colocalising cell. c) Bar plot showing total number of TH+ cells colocalising with EBF3 is similar between mutants and controls (TrkbPenk-WT, 80.5 ± 7.3 vs TrkbPenk-KO, 84 ± 4.0; p = 0.7). d) Bar plot showing the total number of TH+ positive cells that do not colocalize with EBF3 (TrkbPenk-WT, 66.5 ± 5.3 vs TrkbPenk-KO, 71 ± 5.0; p = 0.6). Values are means ± SEM; n = 3 for each genotype/age, male mice. p statistic from unpaired, two-tailed, Student’s t-test. Scale bars, a and b, 100μm; PAG, periacqueductal gray; RFP, retroparafascicular nucleus; fr, fasciculus retroflexus, Related to Fig. 5.
