Fig. 5: LncDACH1 promotes proliferation, migration and phenotype switching during VSMC dedifferentiation by binding to SRPK1. | Nature Communications

Fig. 5: LncDACH1 promotes proliferation, migration and phenotype switching during VSMC dedifferentiation by binding to SRPK1.

From: LncRNA-LncDACH1 mediated phenotypic switching of smooth muscle cells during neointimal hyperplasia in male arteriovenous fistulas

Fig. 5

a RNA Pull Down-LC/MS and Western Blot experiments were used to identify SRPK1 as one of the proteins pulled down by LncDACH1. b RIP assay was used to determine that LncDACH1 was pulled down by anti-SRPK1 antibody (n = 4). c Construction of LncDACH1 fragment (LncDACH1-A-F). fragment A, 0-2085 nt; fragment B, 323-2085 nt; fragment C, 774-2085 nt; fragment D, 1251–2085 nt; fragment E, 774-1251 nt; fragment F, 774-1251[mut] nt. d RNA Pull Down and Western Blot assays were used to determine the binding of LncDACH1 fragments A-E to SRPK1 (n = 3). e RNA Pull Down and Western Blot assays were used to determine the binding of LncDACH1 fragment F to SRPK1 (n = 3). f, g The protein expression levels of SRPK1, p-AKT and AKT after overexpression (n = 3) or silencing (n = 5) of LncDACH1 in VSMC were detected using Western Blot. VSMC is divided into the following four groups: si-NC, si-LncDACH1, si-SRPK1, si-LncDACH1+si-SRPK1. h p-AKT and AKT protein expression levels were detected using Western Blot (n = 3). i, j The proliferation capacity of VSMC was tested by EdU staining assay (n = 3). Scale bar, 25 μm. k The proliferation capacity of VSMC was tested by CCK8 assay (n = 4). l The ability of VSMC to migrate was tested by Wound Healing assays (n = 3). Scale bar, 100 μm. m The ability of VSMC to migrate was tested by Transwell assay (n = 3). Scale bar, 50 μm. n Protein expression levels of VSMC differentiation phenotype markers and dedifferentiation phenotype markers were measured by Western Blot assay (n = 3). PDGF-BB (P), control (CTRL), negative control (NC), overexpression (OE), small interfering (SI). The n numbers represent biologically independent samples. Data are presented as mean values ± SD (b, f–h, j–n). P-values were determined by two-sided nonparametric tests (b, g) and two-sided one-way ANOVA (f, h, j–n) by Bonferroni’s multiple comparisons test. Source data are provided as a Source Data file.

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