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Fig. 3 | Journal of Hematology & Oncology

Fig. 3

From: STOML2 potentiates metastasis of hepatocellular carcinoma by promoting PINK1-mediated mitophagy and regulates sensitivity to lenvatinib

Fig. 3

STOML2 promotes mitophagy in HCC cells under stress. a Expression of top 500 differentially expressed genes upregulated in HCC patients with different STOML2 expression and gene ontology term enrichment analysis for different biological processes controlled by differentially expressed genes among patients with high STOML2 expression. b The autophagic vacuoles were labeled with MDC and then detected by flow cytometry in SMMC-7721 and HCCLM3 cells. The fluorescence intensity of cells represented the autophagic level. c Representative TEM images depicted ultrastructure in SMMC-7721-pCDH or -STOML2-Flag and HCCLM3-shNC- or -shSTOML2. Red arrows indicated autophagic vacuoles. (Scale bars: 500 nm) d Under the treatment of CCCP (10 μM), total protein levels of p62, LC3B I/II and mitochondrial protein VDAC1, Tim23, COXIV were analyzed by Western blot. GAPDH was used as a loading control (left panel). Protein levels of STOML2, p62, and LC3B I/II in mitochondria were examined by purifying mitochondria from HCC cells. COX IV was used as a loading control for mitochondria (middle panel). Quantifying the ratio of LC3B II/LC3B I with image system in total and mitochondrial protein levels (right panel). e, f Confocal microscopy was performed to detect spatial co-localization of mitochondrial protein TOMM20 (red) with LC3B (green) (e, left and middle panel) and LAMP1 (green) (f) in SMMC-7721 and HCCLM3 control and derived cells under the treatment of CCCP (10 μM) for 4 h. (Scale bars:10 μm) The relative fluorescence intensity of LC3B are shown (e, right panel). *P < 0.05; **P < 0.01; ***P < 0.001; ns no significance, WCL whole cell lysate, MITO mitochondria

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