Fig. 6: Dual role of PDGF in regulating FLC differentiation.
From: Tmsb10 triggers fetal Leydig differentiation by suppressing the RAS/ERK pathway

a A schematic summary of signal pathways regulating FLC differentiation and in which Tmsb10 acts as a suppressor. b W-EGFP cells were cultured in the presence (+) or absence (−) of PDGF-AA. Whole-cell extracts prepared from the cells were subjected to western blotting for phospho-AKT (pAKT) and AKT. Representative images of the blots are shown. The positions of molecular weight markers are indicated at the left. c Signal intensities in the blots above were quantified as described in the Materials and Methods. The amounts of pAKT relative to AKT are shown. n = 3. *p < 0.05. d W-EGFP cells were treated with siCnt, siT10, siAkt, or siPten, and then used for testis reconstruction. The reconstructed testes were cultured in the presence of SAG and PDGF for 21 days. The EGFP-positive cells in the reconstructed testes were quantified. n = 3. p < 0.001. e W-EGFP cells were cultured under the same conditions as above. Expression of Gli1 in the cells was determined by qRT-PCR. The data were normalized by Rn18s and are presented as means ± SEM. n = 3. p < 0.001. Letters a, b, and c on the bars in d and e denote significant differences. f W-EGFP cells were cultured under the same conditions as above. Ciliogenesis in the W-EGFP cells was examined by immunostaining for ARL13B. n = 5. *p < 0.001. g W-EGFP cells were cultured in the presence (+) or absence (−) of wortmannin (Wort). Whole-cell extracts were subjected to western blotting for pAKT, AKT, pERK, and ERK. h Signal intensities in the blots above were quantified as described in the Materials and Methods. The amounts of pAKT relative to AKT (left) and pERK relative to ERK (right) are shown. n = 3. **p < 0.01. ***p < 0.001. i W-EGFP cells treated with siRNA for siPten and siCnt. Whole-cell extracts prepared from the cells were subjected to western blotting for pAKT, AKT, pERK, and ERK. Full blot images for b, g, and i are shown in Supplemental Fig. 9. j Signal intensities in the blots above were quantified. The amounts of pAKT relative to AKT (left) and pERK relative to ERK (right) are shown. n = 3. *p < 0.05. **p < 0.01.