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. 2019 Dec 7;11(12):1967. doi: 10.3390/cancers11121967

Figure 3.

Figure 3

PANX1 channel permeability inhibition reduces cell viability and induces cell cycle arrest in MDA-MB-231 and MCF-7 breast cancer cell lines. (A) mRNA levels of PANX1 in MCF-7 cells. qRT-PCR of PANX1, normalized to glyceraldehyde-3-phosphate dehydrogenase GAPDH, in cells treated with 1 mM PBN for 72 h. (B) Analysis of PANX1 protein levels using Western blotting in MCF-7 cells treated with 1 mM PBN for 72 h. Lower panel is a densitometry quantification of the PANX1 and GAPDH bands using Image Lab software. Values represent the average fold change in PANX1 expression, normalized to GAPDH, and relative to control, for a total of three Western blots. (C) Cell viability of MCF-7 cells treated with PBN was assessed by trypan blue dye exclusion assay. Cells were treated with 0.5 or 1 mM PBN for 24, 48 or 72 h. Average cell viability of three independent experiments is displayed as percentage of control (Details of whole blot can be found at Figure S3 and Table S1). (D) Cell cycle distribution analysis was performed by flow cytometry for MCF-7 cells treated with 0.5 mM and 1 mM PBN for 72 h and stained with propidium iodide for cell cycle analysis. Histogram displays averages from three independent experiments. (E,F) Same as panels A and B, but for MDA-MB-231 cells treated with 1 mM PBN for 72 h. (G) Same as (C) but for MDA-MB-231 cells treated with 0.5 or 1 mM PBN for 24, 48 or 72 h. (H) Same as in (D) but MDA-MB-231 cells were used instead. Control denotes DMSO vehicle treated cells. * p < 0.05, ** p < 0.01 and *** p < 0.001.