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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: Gasdermin B over-expression modulates HER2-targeted therapy resistance by inducing protective autophagy through Rab7 activation

Fig. 2

GSDMB-high cells show an increased autophagic flux in response to lapatinib. A-B GSDMB and LC3B protein levels in shNTC, shGB1 and shGB2 HCC1954 (A) and OE19 (B) cells treated with lapatinib (Lap, 2 µM and 0.7 µM, respectively) and/or CQ (10 µM and 50 µM, respectively) for 72 h. Quantification of the relative LC3B-II expression was conducted as described before [34, 35]. C Representative transmission electron microscopy images of shNTC and shGB2 HCC1954 cells treated with the treatment regimens indicated in (A). Quantification of the relative volume density of autophagic vacuoles is shown on the right. At least 25 cells were analyzed per experimental condition. D-E Western blot analysis of GSDMB and LC3B (left panels) in HCC1954 LR (D) and OE19 LR (E) cells and their respective controls (C) treated with or without CQ (10 µM and 50 µM, respectively) for 72 h. LC3B expression (green) analysis by confocal microscopy (right panels) in HCC1954 LR (D) and OE19 LR (E) cells and their controls (C) treated with or without CQ at the concentrations indicated in (A-B). Representative confocal microscopy images were shown, scale bar, 10 µm. Nuclei were counterstained with DAPI. F GSDMB and LC3B protein levels in GSDMB-siRNA-silenced HCC1954 LR cells treated with or without 10 µM CQ for 72 h. Quantification of LC3B-II expression (showed on the right of panels, A-B, D-F) was carried out by densitometric scanning and normalized to GAPDH expression following previous methods [34, 35]. Statistical significance was determined by two-tailed unpaired t-test (*P < 0.05; **P < 0.01). Data are shown as the mean ± s.e.m. Three independent experiments with similar results were performed. NTC, non-targeting control. LR, Lapatinib resistant cells. CQ, chloroquine. Lap, lapatinib

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