Journal: Journal of Biological Chemistry

Article Title: MicroRNA-223-3p inhibits vascular calcification and the osteogenic switch of vascular smooth muscle cells

doi: 10.1016/j.jbc.2021.100483

Figure Lengend Snippet: Figure 6. STAT3 is a potential miR-223-3p target in vascular calcification. A, MiR-223-3p target sites (in red letters) in the STAT3 30 UTRs. B, STAT3 expression in primary vascular smooth muscle cells (mVSMCs) transfected with miR-223-3p or a control mimic (n = 3 per group). C–E, representative western blot and quantification of STAT3 expression in mVSMCs (C), MOVAS (D), and 3T3 cells (E) transfected with miR-223-3p or a control mimic (n = 3 per group). F–G, Representative images (E) and quantification (F) of STAT3 IHC in the brachiocephalic trunks of aged ApoE KO and DKO mice (n = 6 per group). Data are expressed as the mean ± SD. *p <0.05 by unpaired two-tailed Student’s t-test.

Article Snippet: Inhibition of STAT3 expression by siRNA The siRNA against mouse STAT3 and scrambled siRNA were purchased from Santa Cruz Biotechnology, Inc (TX, USA).

Techniques: Expressing, Transfection, Control, Western Blot, Two Tailed Test

Journal: Journal of Biological Chemistry

Article Title: MicroRNA-223-3p inhibits vascular calcification and the osteogenic switch of vascular smooth muscle cells

doi: 10.1016/j.jbc.2021.100483

Figure Lengend Snippet: Figure 7. IL-6-induced VSMC calcification is blocked by miR-223-3p. A, RT-PCR analysis of osteogenic genes in VSMCs. VSMCs were cultured in oste- ogenic media (OIM) with 50 ng/ml mIL-6/sIL-6R complex or PBS for 10 days (n = 3). B, effects of miR-223-3p on osteogenic gene expression in VSMCs. VSMCs were transfected with 100 nM miR-223-3p or a control mimic and cultured in osteogenic medium (n = 3). C, representative western blot and quantification of STAT3 expression in VSMCs cultured as in B for 14 days (n = 4 per group). D, representative images of Alizarin Red S staining of VSMCs cultured as in B for 21 days. E, total calcium content in VSMCs cultured as in B for 14 days (n = 3). F, representative western blot of STAT3 in VSMCs transfected with vector (Lenti-Ctrl) or lentivirus packaged with STAT3(Lenti-Stat3). G, representative images of ARS staining of VSMCs. VSMCs transfected with Lenti-Ctrl or Lenti- Stat3 were cultured in osteogenic media with 50 ng/ml mIL-6/sIL-6R complex for 21 days, and miR-223-3p or a control mimic(100 nM) was transfected in indicated wells. H, total calcium content in VSMCs cultured as in G for 14 days (n = 3). Data are expressed as the mean ± SD. *p <0.05, **p <0.01 compared with the first group, #p <0.05 compared with the second group. Two groups were assessed by unpaired two-tailed Student’s t-test. For comparisons of more than two groups, one-way analysis of variance was used.

Article Snippet: Inhibition of STAT3 expression by siRNA The siRNA against mouse STAT3 and scrambled siRNA were purchased from Santa Cruz Biotechnology, Inc (TX, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Cell Culture, Gene Expression, Transfection, Control, Western Blot, Expressing, Staining, Plasmid Preparation, Two Tailed Test