Fig 2: Effects of SMCs on gap junction proteins expression in HL-1 cells. Cx43, Cx40 and Cx45 were detected using western blot (a), and the relative protein levels of these molecules were determined by densitometric analysis (b–d) (n = 5). Data are shown as mean ± SD. *, P < 0.05. Cx43 connexin43, Cx40 connexin40, Cx45 connexin45, HL-1 HL-1 cells, HL-1/SMC HL-1 cells which were cocultured with contractile SMCs for 48 h, HL-1/SMC TGFß1 HL-1 cells which were cocultured with synthetic SMCs for 48 h. The number of observations (n) represents the number of independent cell preparations

Fig 3: CBX administration decreases the percentages of Cx40-expressing CD4+ and CD8+ T cells in lung tissues of MCT-treated rats. (A) Representative dot plots of CD4+Cx40+ or CD8+Cx40+ double-positive monocytes in lungs. (B) Quantitative analysis of the percentages of CD4+Cx40+ or CD8+Cx40+ T cells indicated that the percentages of CD4+Cx40+ and CD8+Cx40+ T cells were significantly increased in the MCT-treated rats compared with the control group. Compared with the MCT-treated group, the frequencies of CD4+Cx40+ and CD8+Cx40+ T cells were significantly decreased in the MCT + CBX group. The frequencies of CD4+Cx40+ and CD8+Cx40+ T cells were not significantly different between the lungs of control rats and the rats treated with a single intraperito-neal injection of CBX Data are presented as the mean ± standard error of the mean of 6 rats/group. **P<0.01 vs. the control; #P<0.05 vs. MCT-treated rats. APC, allophycocyanin; CBX, carbenoxolone; CON, control; Cx, connexin; CD, cluster of differentiation; FITC, fluorescein isothiocyanate; MCT, monocrotaline.

Fig 4: ß-estradiol supplement reduces the percentages of Cx40- and Cx43-expressing CD4+ and CD8+ T cells in SHR. (A) Representative dot plot and (B) collated data presenting percentages of Cx40- and Cx43-positive CD4+ and CD8+ T cells in each group. Data are presented as the mean ± standard error of three independent experiments (n=6/group). *P<0.05 and **P<0.01, WKY group vs. SHR group without any drug treatment; #P<0.05, SHR group without any drug treatment vs. SHR group with ß-estradiol treatment. Cx, connexin; SHR, spontaneously hypertensive rat; WKY, Wistar-Kyoto rats; SHR + E, SHR + ß-estradiol; CD, cluster of differentiation; FITC, fluorescein isothiocyanate; PE, phycoerythrin; APC, allophycocyanin.

Fig 5: ß-estradiol pre-treatment inhibits Con A-induced protein expression of Cx40 and Cx43 in peripheral blood lymphocytes. The immunoblot analysis of (A) Cx40 and (B) Cx43 in peripheral blood lymphocytes treated with Con A and Con A + ß-estradiol. The bar graph presents the densitometric analysis of relative expression of (A) Cx40 and (B) Cx43 normalized to ß-actin level. Results are presented as the mean ± standard error of five samples/group. *P<0.05 and **P<0.01, control group vs. Con A-stimulated group; #P<0.05, Con A-stimulated group vs. ß-estradiol treated group. Each figure is representative of three independent experiments. Con A, concanavalin A; Cx, connexin.