miR-599 suppressed VSMCs migration.

<p>Overexpression of miR-599 suppressed VSMCs migration. ***p<0.001.</p

TGFb2 involved in the effect of miR-599 in VSMCs.

<p>(A) The protein expression of TGFb2 was measured by western blot. (B) Overexpression of TGFb2 promoted miR-599-induced inhibition of VSMCs proliferation. (C) Ectopic expression of TGFb2 promoted the expression of type I collagen, in miR-599 overexpressing VSMCs. (D) The mRNA expression of type V collagen in VSMCs was measure by qRT-PCR. (E) The mRNA expression of proteoglycan in VSMCs was measure by qRT-PCR. *p<0.05, **p<0.01 and ***p<0.001.</p

miR-599 inhibited matrix gene expression in VSMCs.

<p>(A) Overexpression of miR-599 suppressed the type I collagen mRNA expression. (B) The protein expression of type I collagen in VSMCs was measure by western blot. (C) Overexpression of miR-599 suppressed the type V collagen mRNA expression. (D) The protein expression of type V collagen in VSMCs was measure by western blot. (E) Overexpression of miR-599 suppressed the proteoglycan mRNA expression. (D) The protein expression of proteoglycan in VSMCs was measure by western blot.</p

miR-599 suppressed VSMCs proliferation.

<p>(A) The expression of miR-599 was measured by qRT-PCR. (B) CCK-8 analysis showed that overexpression of miR-599 suppressed VSMCs proliferation. (C) The mRNA expression of PCNA in VSMCs was measure by qRT-PCR. (D) The protein expression of PCNA in VSMCs was measure by western blot. (E) The mRNA expression of ki-67 in VSMCs was measure by qRT-PCR. (F) The protein expression of ki-67 in VSMCs was measure by western blot.*p<0.05 and ***p<0.001.</p

miR-599 is inhibited by PDGF-bb in VSMCs.

<p>(A) PDGF-bb can induce VSMCs proliferation at time-dependently. (B) PDGF-bb can induce VSMCs proliferation at dose-dependently. (C) The expression of miR-599 was measured by qRT-PCR. (D) The expression of miR-599 was measured by qRT-PCR. *p<0.05, **p<0.01 and ***p<0.001.</p

TGFb2 is a direct target of miR-599 in VSMCs.

<p>(A) There is a potential seed sequence of miR-599 in the 3’UTR of TGFb2. (B) miR-599 inhibited the luciferase activity in the TGFb2 with wild-type 3’UTR, whereas, luciferase activity was not drop in the mutant binding sites 3’UTR of TGFb2. (C) Overexpression of miR-599 suppressed the protein expression of TGFb2. ***p<0.001.</p