Fig 1: Berberine improves the impaired autophagic activity in EPCs isolated from VLU patients. (A) The expression of autophagy-related protein LC3B was detected by immunofluorescence staining in the normal EPCs, untreated EPCs isolated from VLU patients and berberine-treated EPCs isolated from VLU patients. (B) The protein expression of RRAGB, p-mTOR, mTOR and LC3A/B in the normal EPCs, untreated EPCs isolated from VLU patients and berberine-treated EPCs isolated from VLU patients was detected by western blotting. ∗∗p < 0.01 vs. Control; ##p < 0.01.
Fig 2: MiR-21-3p directly targets RRAGB whose overexpression inhibits the proliferation, migration and tubular formation of EPCs isolated from VLU patients. (A) The protein expression of RRAGB in the model EPCs transfected with miR-21-3p mimics was detected by western blotting. ∗∗p < 0.01 vs. Model + miR-21-3p NC. (B) The binding correlation between miR-21-3p and RRAGB was identified by the luciferase reporter assay. ∗∗p < 0.01 vs. pmirGLO + miR-21-3p agomir NC; ##p < 0.01. (C) The proliferative ability of model EPCs transfected with RRAGB overexpression plasmid (RRAGB-OE) was analyzed by CCK-8 assay. ∗∗p < 0.01 vs. Model + NC. (D) The migration ability of model EPCs transfected with RRAGB-OE was detected by Transwell migration assay. ∗∗p < 0.01 vs. Model + NC. (E) The tube-forming capability of model EPCs transfected with RRAGB-OE was evaluated by tubule formation assay. ∗∗p < 0.01 vs. Model + NC.
Fig 3: RRAGB overexpression upregulates the protein expression of mTOR pathway-related proteins (p-mTOR and mTOR) while downregulates the autophagy-related proteins (LC3A/B and LC3B). (A) The protein expression of LC3A/B, p-mTOR and mTOR in the model EPCs transfected with RRAGB-OE was detected by western blotting. (B) The expression of autophagy-related protein LC3B was detected by immunofluorescence staining in the model EPCs. ∗∗p < 0.01 vs. Model + NC.
Supplier Page from Abcam for Recombinant Human RRAGB protein (GST tag N-Terminus)