Hypoxia Promotes Vascular Smooth Muscle Cell (VSMC) Differentiation of Adipose-Derived Stem Cell (ADSC) by Regulating Mettl3 and Paracrine Factors

Figure 3

Effect of Mettl3 knockdown on the VSMC differentiation potential of ADSCs. (a) Fluorescence protein marker was used to determine the transfer efficiency of Mettl3 knockdown in ADSCs. After transfection for 72 h, the cells were observed under a microscope (A). (B) is an immunofluorescence image taken at the same time. The “white” scale bars represent 20 μm (original magnification ×200). (b) The expression level of Mettl3 was determined using Western blotting in the Mettl3-shRNA and Mettl3-shCtrl groups. Vinculin was used as an internal control. The band intensities were analyzed using the ImageJ software. (c) The m6A level of ADSCs in the Mettle3-shaCtrl and Mettl3-sh1 groups. (d) The total RNA level of ADSCs in the Mettle3-shaCtrl and Mettl3-sh1 groups. (e) The mRNA expression level of α-SMA, SM22α, calponin, and SM-MHC in the Mettl3-shRNA and Mettl3-shCtrl groups was assessed using qRT-PCR after 7 and 14 days of induction. GAPDH was used as an internal control. All of the results represent the of three independent experiments (). Significant difference compared with the control ().

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