Figure 2
Loss of Ahr promotes GCP cycle arrest through enhanced activation of the TGF-β/SMAD3 axis. (a) Western blots of total cell lysates of isolated P7 GCPs from control and Ahr cKO cerebella with antibodies specific to phosphorylated S423/S425 residues of SMAD3 and phosphorylated S465/467 residues of SMAD2 proteins as well as antibodies against total SMAD2/3 and GAPDH (loading controls). Molecular weight markers are indicated on the left. (b) Quantification of band optic density for phosphorylated SMAD3, relative to total SMAD2/3, normalized to GAPDH levels. (c) Quantification of %Ki67 positive cells in non-treated (−) and 1 uM SIS3 (+SIS3) treated cultures. (d) Quantification of %Neurod1 positive cells. Data shown in (c,d) is representative of GCPs (on average 100 cells counted from 4 different fields of view from triplicate wells) isolated from 3 animals of each genotype, cultured for 24 hours in triplicate. Data was analyzed by Student’s t test (p < 0.001 (***), p < 0.01 (**), p < 0.05 (*)). Bars represent mean values +/− SEM.
