Fig. 2: PER2-dependent promotion of cuproptosis inhibits OSCC cell proliferation in vitro. | Cell Death & Disease

Fig. 2: PER2-dependent promotion of cuproptosis inhibits OSCC cell proliferation in vitro.

From: PER2 interaction with HSP70 promotes cuproptosis in oral squamous carcinoma cells by decreasing AKT stability

Fig. 2

A RT-qPCR to determine DLAT, PDHB and SLC31A1 mRNA expression in OSCC cells with overexpression of PER2. B Western blotting to determine DLAT, PDHB, SLC31A1, SDHB and DYPD protein expression in OSCC cells with overexpression of PER2. C Copper Colorimetric Assay Kit to determine the concentration of copper in OSCC cells with overexpression of PER2. D Copper affinity chromatography assay to determine the binding of copper to DLAT protein in OSCC cells with overexpression of PER2. E Non-denaturing gel electrophoresis assay to detect DLAT oligomers in OSCC cells with overexpression of PER2. F Immunofluorescence assay to detect DLAT oligomers in OSCC cells with overexpression of PER2 (yellow, DLAT oligomer; green, DLAT; red, Mitotracker; blue, DAPI; scale bars = 50 μm; three independent experiments). G TEM to observe and quantify the number of vacuolated mitochondria in OSCC cells with overexpression of PER2, as well as the observation that mitochondria appeared to be deformed and swollen (red arrows indicate mitochondria; three independent experiments). H Micro-mitochondrial Complex I and II Activity Assay Kit to detect activity of mitochondrial complexes I and II in OSCC cells with overexpression of PER2. I Fumarate Assay Kit and α-KG Assay Kit to detect the concentration of TCA intermediate metabolites (fumarate and α-ketoglutarate) in OSCC cells with overexpression of PER2. J CCK-8 assay to determine levels of cell proliferation in OSCC cells with overexpression of PER2. K MTT assay to determine levels of cell proliferation in OSCC cells with overexpression of PER2. L CCK-8 assay showed a significant increase in proliferation level of OE-PER2-SCC25 cells after the addition of copper chelator TTM compared with OE-PER2-SCC25 cells. M MTT assay showed a significant increase in proliferation level of OE-PER2-SCC25 cells after the addition of copper chelator TTM compared with OE-PER2-SCC25 cells. All data represent three replicate independent experiments. Data are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

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