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. 2021 Dec 6;18(8):1822–1840. doi: 10.1080/15548627.2021.2002108

Figure 3.

Figure 3.

SH3BGRL promotes autophagy flux in breast cancer cells.(A-C) Immunoblots of SQSTM1, LC3B-I and LC3B-II in indicated cells with or without Dox treatment (A). ACTB was used as an internal loading control. Statistical analyses of SQSTM1 expression (B) and LC3B-II:LC3B-I ratio(C) in indicated cells. ***P < 0.001, ****P < 0.0001, n.s., no significance. (D) Immunoblots of SQSTM1, LC3B-I and LC3B-II in indicated cells. Cells were pretreated with 50 nM Baf A1 for 4 h and cultured in either normal culture medium (upper panel, Normal) or EBSS medium (lower panel, EBSS) for another 2 h, respectively. ACTB was used as an internal loading control. (E) Immunoblots of autophagy-related proteins and the cleaved c-CASP3 in MCF-7 SH3BGRL overexpressing cells or MDA-MB-453 SH3BGRL knockdown cells. Cells were treated with Dox along with or without 50 nM BafA1 for 4 h. (F,G) Representative immunofluorescence staining of LC3B puncta in indicated cells with Serum-free medium (F) for 12 h or EBSS culture for 6 h (G). Cells were counterstained with DAPI in blue. Bars: 25 μm. (H) Quantifications of LC3B puncta intensity in the assayed cells (F,G) were presented as histograms. ***P < 0.001, ****P < 0.0001.