SM22α Loss Contributes to Apoptosis of Vascular Smooth Muscle Cells via Macrophage-Derived circRasGEF1B

Figure 3

Macrophage-derived circRasGEF1B induces VSMC apoptosis. (a, b) Flow cytometry analysis (a) or Western blot (b) of cell apoptosis in MOVAS cells, a mouse VSMC line, pretreated with the CM from saline- or LPS-stimulated RAW264.7 cells. (c, d) Flow cytometry analysis of cell apoptosis in MOVAS cells pretreated with IgG or TNF-α neutralizing antibody (c) or in WT and Sm22α^-/- VSMCs (d) treated with Ang II for 24 h. (e–g) qRT-PCR of lincRNA-Cox2, miR-146a, miR-155, circRasGEF1B, circRNA-010231, circRNA-010056, and circRNA-003780 in RAW264.7 cells (e), in the conditional media from RAW264.7 cells (f), and in MOVAS cells pretreated with the CM from RAW264.7 cells (g). (h) qRT-PCR of circRasGEF1B in RAW264.7 and MOVAS cells. (i) Confocal FISH images of circRasGEF1B in VSMCs pretreated with CM from RAW264.7 cell or not. Bars: 10 μm. (j) RT-PCR of circRasGEF1B in MOVAS cells transfected with the pLCDH empty vector or pLCDH-circRasGEF1B plasmid. (k, l) TUNEL assay (k) and Western blot (l) of Bax and Bcl-2 in MOVAS cells transfected with the pLCDH empty vector or pLCDH-circRasGEF1B plasmid. Bars: 100 μm (k). (m) TUNEL assay of cell apoptosis in WT or Sm22α^-/- VSMCs transfected with the pLCDH-circRasGEF1B plasmid. Bars: 100 μm. Data are presented as of three independent experiments. , , and .

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