Figure 2.

Effect of connexin and pannexin hemichannels blockers on the generation of Ca²⁺ signals in white adipocytes due to the application of Ca²⁺-free medium. (A,B) Effects of connexin (Carbenoxolone (CBX), 100 µM; Octanol, 1 mM; or Gap26, 100 µM) and pannexin (Probenecid (PROB) 1 mM) hemichannels blockers on the Ca²⁺ oscillation (A) and Ca²⁺ transients (B) of white adipocytes upon application of Ca²⁺-free medium. Between the first and second application of the Ca²⁺-free medium there was a 30 min pause in the Ca²⁺-dynamics registration (recording). (C) Cell knockdown of the Cx43 hemichannels completely suppresses Ca²⁺ oscillations and significantly suppresses the amplitude of Ca²⁺ transients in white adipocytes upon application of a Ca²⁺-free medium. NE—application of 1 µM norepinephrine, an adrenergic receptor agonist. (D) Effect of the investigated blockers on the amplitude of the Ca²⁺ oscillations and transients upon repeated Ca²⁺-free application. The amplitude of the Ca²⁺ response to the first application of the Ca²⁺-free medium is taken as 100%. Abbreviations: CBX—Carbenoxolone, 100 µM; Octanol, 1 mM; Proadifen, 100 µM; Gap26, 100 µM; blockers of connexin hemichannels: ¹⁰Panx, 100 µM and Probenecid, 1 mM; and blockers of pannexin hemichannels, Cx43-KD; Panx-KD—Cx43 and Panx1 knockdown using Gja1 siRNA and Panx1 siRNA; Scrambled—cells transfected with an siRNA whose sequence differed from that of the siRNA against Cx43 and Panx1; 2 response—the amplitude of Ca²⁺ responses to the Ca²⁺-free medium application without inhibitors. The differences between the experimental groups and control are marked with asterisks: ** p < 0.01, *** p < 0.001; n/s—no significant difference.