MicroRNA-93 Regulates Hypoxia-Induced Autophagy by Targeting ULK1

Figure 2

miR-93 inhibits hypoxia-induced autophagy. (a) The expression of ULK1 is induced by hypoxia. MEFs are subjected to hypoxia conditions for 0 h, 6 h, 12 h, and 24 h. Endogenous ULK1, P62, LC3, and β-actin were blotted by corresponding antibodies. Densitometric ratios of LC3-II/I from the samples were quantified by using ImageJ. (b) Expression levels of endogenous miR-93 and ULK1 in MEFs under hypoxia condition for 0 h, 6 h, 12 h, or 24 h by qPCR. (c) MEFs were transfected with scramble NC (NC), miR-93 mimics (93), scramble IN-NC (IN-NC), or miR-93 inhibitor (IN-93) for 12 h; afterward, MEFs were sealed in a hypoxic or normoxic condition for another 24 h with or without Bafilomycin A1 (BAF1). Samples were blotted with anti-ULK1, anti-P62, anti-LC3, and anti-β-actin antibodies. (d) MEFs were treated as the same as (c). Cells were fixed and immunostained with anti-ULK1 (green) and anti-P62 (red) antibodies. Scale bar, 20 μm.

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