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. 2020 Jul 13;11(7):526. doi: 10.1038/s41419-020-02745-x

Fig. 4. Circ-ADAM9 regulates apoptosis, autophagy, and angiogenic function of EPCs induced by high glucose.

Fig. 4

EPC dysfunction was induced by high glucose (30 mM) treatment for 24 h. Mannitol was used as an osmolar control treatment. Cells were transfected with circ-ADAM9 knockout (sh-circ-ADAM9) or overexpression (LV-circ-ADAM9) lentiviral construct or negative control lentivirus (sh-NC or EV-circ-ADAM9). a Flow-cytometric analysis of apoptosis using annexin V/PI. Annexin V+/PI+ or Annexin V+/PI– (quadrants 2 and 3) cells were defined as apoptotic cells. b Protein levels of cleaved-CASP3, BAX, and BCL-2 as detected by western blotting. c Representative images showing LC3 staining in different groups of EPCs infected with GFP-RFP-LC3 adenovirus for 24 h. Scale bar: 20 μm. d Western blot analysis of LC3B-II/LC3B-I and SQSTM1/p62 levels. e The angiogenic capability of EPCs was determined by a tube formation assay. Tube length was normalized to that in the control group. Scale: 200 μm. f Protein levels of VEGF as detected by western blotting. *P < 0.05, **P < 0.01, n = 3.