Journal: BMC immunology

Article Title: miR32-5p promoted vascular smooth muscle cell calcification by upregulating TNFα in the microenvironment.

doi: 10.1186/s12865-019-0324-x

Figure Lengend Snippet: Fig. 2 The influence of TNFα expressed by BV2 cells for VSMC calcification. a-f Alizarin red staining analysis of VSMC calcification in differently stimulated condition (40×), a group1 added 10% BV2 culture supernatant and 10 mM β-glycerophosphate, b group3 added 10 mM β- glycerophosphate and 10 ng/ml mouse TNFα, c group5 10 mM β-glycerophosphate, d group2 added 10% BV2 culture supernatant, which was incubated with rabbit anti-mouse TNFα antibody, and 10 mM β-glycerophosphate, e group4 added 10 ng/ml mouse TNFα, f group6 added equal volume PBS. g-i qRT-PCR analysis for the expression of Runx2 and α-SMA in VSMC, g the mRNA expression of Runx2 and α-SMA in groups 1 and 2, h the mRNA expression of Runx2 and α-SMA in groups 3 and 4, i the mRNA expression of Runx2 and α-SMA in groups 5 and 6. Bars represent mean ± S.D. (n ≥3). Significant difference marked with one (p < 0.05) or two (p < 0.01) asterisks

Article Snippet: According to the report by Ma et al [19] with some modification, the VSMCs were divided into 6 groups, cultured in 12-well plates with 1ml DMEM with 10% FBS and 100 U/ml penicillin-streptomycin, and treated as follows: group 1 received 10% BV2 culture supernatant (treated with LPS) and 10mM β-glycerophosphate (Sigma, Poole, Dorset, UK); group 2 received 10% BV2 culture supernatant (treated with LPS), which was incubated with 7 μl rabbit anti-mouse TNFα antibody (BOSTER Biological Technology Co. Ltd., China) at 4 °C for 12 h and 10mM β-glycerophosphate; group 3 received 10mM βglycerophosphate and 10 ng/ml mouse TNFα (BOSTER Biological Technology Co. ltd., China); group 4 received 10 ng/ml mouse TNFα; group 5 received 10mM βglycerophosphate; and group 6 received equal volume of PBS.

Techniques: Staining, Incubation, Quantitative RT-PCR, Expressing