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Fig. 4 | BMC Cancer

Fig. 4

From: Molecular features of the cytotoxicity of an NHE inhibitor: Evidence of mitochondrial alterations, ROS overproduction and DNA damage

Fig. 4

Activation of autophagy by HMA. Silencing of ATG7 and Beclin-1 by siRNA in HCT-116 cells. Impact of HMA on Ubiquitin and GRP78. a Effect of silencing of ATG7 and Beclin-1 on HCT-116 cell survival after a 30-μM HMA treatment (for 24 h). Cell death was monitored by flow cytometry after Annexin V/PI staining; four fractions can be distinguished: PI+/A+: late apoptotic/necrotic cells; A+: early apoptotic cells; PI+: necrotic cells; unstained: alive cells. b Western blot analysis of Atg7 and Beclin-1 in HCT-116 cells after transfection with siRNAs; γ-tubulin: loading control. Red rectangles refer to the siRNA used in a). c Immunofluorescence analysis of p62 (green fluorescence) in HCT-116 cells untreated and treated with 20 μM and 40 μM HMA for 24 h. Nuclei were counterstained with DAPI (blue fluorescence). Scale bar: 50 μm. A representative experiment out of three is shown. d Immunofluorescence analysis of Ubiquitin and GRP78 (red fluorescence) in HCT-116 cells untreated and treated with 20 μM HMA for 24 h; nuclei were counterstained with DAPI (blue fluorescence)

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