Journal: International Journal of Biological Sciences
Article Title: Acute Exposure to Cadmium Triggers NCOA4-Mediated Ferritinophagy and Ferroptosis in Never-Smokers Oral Cancer Cells
doi: 10.7150/ijbs.111228
Figure Lengend Snippet: Effects of CdCl 2 exposure in CAL27T cells. To obtain CAL27T (CAL27 Tolerant to Cd) CAL27 were exposed to low doses of CdCl 2 (10μM) for 30 days. (A) PI flow cytometry assay and relative histograms of CAL27T cells treated with CdCl 2 (26.01μM for 12h). % of dead cells (PI positive) are reported in each dot plot. (B) Quantification of iron and cadmium intracellular amount through ICP-MS in CAL27T cells treated with 26.01μM CdCl 2 (12h). (C) Western blot analysis and relative optical densitometry of NCOA4, FtH1, LC3B (I-II), mTORC1 and p-mTORC1 in CAL27T cells treated with 26.01μM CdCl 2 (12h). GAPDH was used as normalization control for protein quantification. (D) Fluorescence microscopy analysis of LIP content with FerroOrange dye in CAL27T cells after treatment with 26.01μM CdCl 2 (12h). Flow cytometry analyses and relative histograms of mitochondrial ROS amount (E) , mitochondrial membrane potential (F) and lipid peroxidation (G) assessed by using MitoSOX, TMRM and BODIPY-C11 reagents, respectively, in CAL27T cells following treatment with 26.01μM CdCl 2 (12h). Each experiment was performed in triplicate. Histograms are presented as mean ± SD. p -value: *≤0.05; ***≤0.001; ****≤0.0001. ns: not significant.
Article Snippet: Human oral squamous cell lines (OSCC) - CAL27, OT1109, SCC090, and SCC154 - were purchased from the American Type Culture Collection (ATCC, Rockville, MD, United States).
Techniques: Flow Cytometry, Western Blot, Control, Fluorescence, Microscopy, Membrane