Figure 5 | Scientific Reports

Figure 5

From: Cardiac inflammatory CD11b/c cells exert a protective role in hypertrophied cardiomyocyte by promoting TNFR2- and Orai3- dependent signaling

Figure 5

Conditioned medium from CD11b/c cells isolated from EACH heart activates Ca2+ influx in a manner sensitive to TNFR2-Ab and Orai inhibitor YM58483. (A) Schematic representation of the protocol where rats implanted with an iso-pump for fourteen days were subjected to echographic analyses and developed EACH. (B) CD11b/c cells, cardiomyocytes and fibroblasts were isolated from EACH hearts and cultured for 18 hours before recovery and concentration of conditioned media. CD11b/c cells pre-incubated with/without the anti-inflammatory drug, semapimod, prior in vitro LPS application to induce pro-inflammatory activation, were isolated from normal hearts and cultured for 18 hours before recovery and concentration of conditioned media. Iso-treated hypertrophied cardiomyocytes were loaded with Fura2 before measurement of voltage- and store-independent Ca2+ influx, 1st in the absence and 2nd in the presence of these conditioned media (Cmed). (C) Only Cmed from CD11b/c cells isolated from EACH hearts activates a voltage- and store-independent Ca2+ influx (D) in a manner sensitive to TNFR2-Ab and Orai inhibitor YM58483. (E) Cmed from CD11b/c cells isolated from normal hearts and in vitro stimulated with LPS activates a voltage- and store-independent Ca2+ influx in a manner sensitive to neutralizing TNFR2-Ab, Orai inhibitor YM58483 and anti-inflammatory pretreatment with semapimod. Number of cells analyzed, number of cell isolations (rats) and number of Cmed tested as indicated, mean ± SEM of cells, Wilcoxon matched-paired tests to examine if the mean of the 2nd rate differs from the 1st one, arbitrarily set as 1, ***p < 0.001, ****p < 0.0001.