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. 2015 Nov 6;5:16295. doi: 10.1038/srep16295

Figure 3. SHED-CM attenuates apoptosis and inflammation in cardiac myocytes.

Figure 3

Rat neonatal cardiac myocytes were treated with control and SHED-CM under 24 h of hypoxia or 48 h of normoxic conditions. (a) Representative photomicrographs of TUNEL-positive cardiac myocytes. Apoptotic nuclei were identified by TUNEL-staining (green) and total nuclei by DAPI counterstaining (blue). (b) Quantitative analysis of TUNEL-positive cardiac myocytes treated with vehicle and SHED-CM under 24 h of hypoxia or 48 h of normoxic conditions. TUNEL-positive cells were counted in three randomly chosen microscopic fields of the three different slides and expressed as a percentage of the total number of nuclei. (c) Cell viability of cardiac myocytes was analyzed by WST-8 assay after 48 h serum deprivation treated with vehicle and SHED-CM. Results are presented as mean ± SEM (n = 3 in each group). (d) Effect of SHED-CM on LPS-induced expression of TNF-α, IL-6, and IL-1β in cardiac myocytes. Cardiac myocytes were pretreated with vehicle, 20% SHED-CM or SHED-CM for 1 h and stimulated with or without LPS (100 ng/ml) for 6 h. The mRNA expression of cytokines was measured by real-time PCR and expressed relative to GAPDH levels (n = 3 in each group). Results are presented as mean ± SEM.