Fig 2: (A) The Western blotting results showed that incubating aortic VSMCs with 10 µM T16Ainh-A01, 100 µM arctigenin, and 30 µM siRNA TMEM16A led to a decrease in ESM1. Data are expressed as mean ± SD, and relative protein expression was quantified using one-way ANOVA and p-value adjusted for multiple comparisons with Sidak multiple-comparison test (n = 3), **** p < 0.0001. (B) Microscopic images showed immunofluorescence staining of ESM1 (green) in thoracic aortas (n = 4); blue represents nucleus. Mean fluorescence was quantified using one-way ANOVA and p-value adjusted for multiple comparisons with Sidak multiple-comparison test (n = 4), *** p < 0.001, **** p < 0.0001. (C) 30uM siESM1 knockdown EMS1 protein for 48 h in the ESM1 knockdown cells compared to the HS group. Data are expressed as mean ± SD, * p < 0.05, t-test (n = 3). (D) ESM1, VCAM-1, and ICAM-1 protein expression dramatically declined after silencing ESM1 in VSMCs. Data are expressed as mean ± SD, and relative protein expression was quantified using one-way ANOVA and p-value adjusted for multiple comparison with Sidak multiple-comparison test (n = 3), * p < 0.05, ** p < 0.01, **** p < 0.0001. (E) Aortic VSMCs were cultured with NS, HS, ARC, siTMEM16A siESM1 for 24 h and T16 for 30 min, and the levels of CXCL16 in the culture media were quantified using the enzyme-linked immunosorbent assay (ELISA). Data are expressed as mean ± SD, and relative CXCL16 expression was quantified using one-way ANOVA and p-value adjusted for multiple comparisons with Sidak multiple-comparison test (n = 3), **** p < 0.0001 (n = 3). NS: normal salt group, HS: high-salt group, T16: HS+ T16Ainh-A01 group, ARC: HS+ arctigenin (100 mg/kg) group, siTMEM16A: HS+siRNA TMEM16A group. siEMS1: HS+ siRNA ESM1 group.