Figure 4
From: The syntaxin-binding protein STXBP5 regulates progerin expression
Stable knockdown of STXBP5 altered the expression of progerin and other senescence-associated genes. (A–D) HA-progerin-expressing HEK293 cells were transfected with one of three STXBP5 guide RNAs (STXBP5 guide1, STXBP5 guide2 or STXBP5 guide3) or a control guide RNA (gAAVS1) and selected with puromycin for 3 days to obtain a cell line with stable knockdown. (A) Western blotting was performed with an anti-HA antibody for HA-tagged progerin expression, with an anti-LaminA/C antibody and with an anti-STXBP5 antibody. \(\beta\)-Actin was used as a loading control. (B) The mRNA expression levels of Lamin A, progerin, Lamin B1, Lamin C, P53, P21, IL6 and IL8 in HA-progerin-expressing HEK293 cells transfected with STXBP5-specific guide RNAs were measured via qRT-PCR. (C) Representative images of SA-\(\beta\)-gal staining from three independent experiments. Scale bars, 50\(\mu\)m. (D) The Lamin B1 expression level was measured woth anti-Lamin B1 antibody, and a graph was generated. (E) Gene-gene interaction network among STXBP5, LMNA and LMNB1. The data are presented as the means ± SEMs. Comparisons were performed with two-way ANOVA (B) or one-way ANOVA (C,D) with the Brown–Forsythe test for multiple comparisons. ns, p > 0.05; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.
