Figure 2 | Scientific Reports

Figure 2

From: Malignant Transformation of Human Bronchial Epithelial Cells Induced by Arsenic through STAT3/miR-301a/SMAD4 Loop

Figure 2

Arsenic induced miR-301a expression dependent on IL-6/STAT3 signaling. (A) BEAS-2B cells were treated with arsenic (5 and 10 μM) for 24 h and with H2O2 as positive control. The levels of IL-6 in the culture medium was determined by using ELISA Kit. (B) BEAS-2B cells were treated with different concentrations of IL-6 (0, 10, 50, and 100 ng/mL) and IL-6 plus Stat3 inhibitor (S31-201, 30 μM), at 6 h and the expression of miR-301a was determined by qPCR. (C) BEAS-2B cells were treated with arsenic (As, 10 μM), arsenic (As) and S31-201, arsenic (As) and IL-6 neutralizing antibody (anti-IL-6) at 6 h and the expression of miR-301a was determined by qPCR. (D) BEAS-2B cells were transfected with Anti-control (Anti-control) and LNA-Anti-miR-301a(Anti-miR-301a). After 48 h post transfection, cells were treated with arsenic (5 μM and 10 μM) for 24 h and western blot analyses of phospho-Stat3 (pStat3), Stat3, IκBα expression. Gel pictures were acquired by using Tanon5200 and the exposure time was 60 sec for pStat3, 20 sec for Stat3, 20 sec for IκBα, and 5 sec for β-actin. (E) Protein expression from (D) were quantified by band intensity and normalized to β-actin. (F) pStat3 expression was quantified by band intensity and normalized to total Stat3. Values represented the mean ± SD of three independent experiments. *P < 0.05 and **P < 0.01 indicate a significant difference between the indicated groups, or between none treatment and treatment groups.

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