Fig. 5: Activation of the RAS/ERK pathway by PDGF. | Communications Biology

Fig. 5: Activation of the RAS/ERK pathway by PDGF.

From: Tmsb10 triggers fetal Leydig differentiation by suppressing the RAS/ERK pathway

Fig. 5

a Reconstructed testes were incubated in the absence (open bar) or presence of PDGF-AA (green bar), SAG (dark blue bar), or both PDGF-AA and SAG (red bar). The EGFP-positive cells in the reconstructed testes were analyzed quantitatively after incubation for 21 days. n = 3. p < 0.001. b W-EGFP cells were cultured under the same conditions as above. Expression of Gli1 in the cells was examined by qRT-PCR. The data were normalized by Rn18s and are presented as means ± SEM. n = 3. p < 0.01. c Whole-cell extracts prepared from W-EGFP cells were cultured in the presence (+) or absence (−) of PDGF-AA and subjected to Western blotting for ERK and pERK. The position of a molecular weight marker is indicated on the left. d Signal intensities of the blots above were quantified as described in the Materials and Methods. The amounts of pERK relative to ERK are shown. Letters a, b, and c on the bars in a, b, and d denote significant differences between the cell groups. n = 3. p < 0.01. e W-EGFP cells were treated with siRNA for siT10 and thereafter cultured in the presence (+) or absence (−) of PDGF-AA. Whole-cell extracts prepared from the cells were subjected to western blotting for ERK and pERK. f Signal intensities of the blots above were quantified. The amounts of pERK relative to ERK are shown. n = 3. ***p < 0.001. Full blot images for c and d are displayed in Supplemental Fig. 9. g A tentative schema for the function of PDGF is shown. It was assumed that another signal pathway could be activated downstream of PDGF. h W-EGFP cells were cultured in the absence (−) or presence (+) of PDGF-AA and SAG. Ciliogenesis in the W-EGFP cells was examined by immunostaining for ARL13B. Ciliated cells were counted and the ratios of these cells to all cells (%) are plotted. n = 3. *p < 0.001.

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