Figure 5.
Podocyte injury and mitotic catastrophe require Sox4 in vitro
(A) Western-blot analysis showed the expression of Sox4, p53, p21cip1/waf1, CyclinB, and cdc2 (n = 3). (B) Flow cytometry analysis illustrated the cell-cycle progression of podocytes (n = 3). (C) Immunostaining of mitosis with antibodies against α-tubulin (green) and H3-Ser10 (red) (n = 3). Scale bar, 10 μm. (D)The percentages of podocytes with normal or abnormal mitotic processes were monitored and analyzed (n = 3). (E) Morphological changes in the podocyte cytoskeleton (n = 3). Scale bar, 50 μm. (F) The expression of Desmin, podocin, synaptopodin, and ZO-1 was measured using western blotting (n = 3). (G–I) Immunofluorescence staining was performed to determine the intensities of podocin (G), Desmin (H), and p-cadherin (I) under HG conditions (n = 4). Scale bar, 50 μm. (J) Cell migration was detected using Transwell assays (n = 5). Scale bar, 100 μm. (K) Flow cytometry revealed the apoptosis rate of podocytes (n = 5). (L) Flow cytometry clarified the proportion of podocytes in G2/M phase (n = 4). (M) Immunostaining illustrated morphological changes associated with normal or abnormal mitosis using antibodies against α-tubulin (green) and H3-Ser10 (red) (n = 3). Scale bar, 10 μm. (N) The percentages of podocytes with normal or abnormal mitotic processes were monitored and analyzed (n = 3). Error bars represent ±SD. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.