Fig. 5

Effect of hypertension-mediated inflammation and blocking of the gap junction on gap junctional intracellular communication (GJIC) between peripheral blood lymphocytes from spontaneously hypertensive rats (SHRs). The X-axis represents the parameter’s signal value in the channel numbers (count) and the Y-axis represents the number of events per channel number (calcein AM- and/or DiIC₁₈-positive cells). a – Control experiments of DiIC₁₈ or calcein AM single-stained lymphocytes and DiIC₁₈–calcein double-stained lymphocytes were performed in top parallel. In the histogram, the red line depicts background fluorescence while the blue line depicts fluorescence in DiIC₁₈ or calcein AM single-stained lymphocytes and DiIC₁₈–calcein double-stained lymphocytes. Isolated peripheral blood lymphocytes from Wistar-Kyoto (WKY) rats and SHRs were stained with calcein AM or DiIC₁₈, and co-cultured for 3 h in the absence or presence of Con A (5 μg/ml) and Gap27 (500 μM) as described in the Materials and Methods section. Direct calcein AM transfer through GJCs from donor lymphocytes (calcein AM single-stained cells) to recipient lymphocytes (DiIC₁₈ single-stained cells) was assessed using flow cytometry. DiIC₁₈–calcein AM double-stained fluorescent cells expressed as a percentage of the total number of peripheral blood lymphocytes. The effect of Gap27 on GJIC of the co-cultured peripheral blood lymphocytes was studied. b – Bar graph of the mean percentages of DiIC₁₈–calcein double-positive cells from three independent experiments ± SEM. ^&&p < 0.01 versus WKY group. *p < 0.05, compared with the control group in the same column (n = 5); ^#p < 0.05, compared with the Con A-stimulated group in the same column (n = 5)