Ouabain and miR-34a Treatment Augments Autophagy in Cancer Cells
(A) Phase contrast images of A549 cells treated with miR-Control, miR-34a, ouabain, and combination of miR-34a (40 nM) with ouabain (IC50, 30 nM). All images are taken at 200× magnification. Images from cell treated with ouabain or miR-34a combination show large vacuoles, consistent with an autophagic phenotype.
(B) Fluorescent images from A549 cells with the RFP-GFP-LC3B reporter assay for autophagy. Autophagosomes (green) and LC3B-positive autophagolysosome (red) and nucleus (blue) are shown with quantification of number of respective positive cells as a percentage of total number of cells (lower panel).
(C) Western blot data showing LC3A/B bands in cells treated with miR-control, ouabain (30 nM), or miR-34a (40 nM) combinations in A549 cells. *p < 0.05, **p < 0.01, ***p < 0.001, Student's t test. Bars and errors bars represent the means and the corresponding SEMs for n ≥ 3.