Figure 4.

Irisin reduced apoptosis during ischemic injury in JEG-3 cells. (A) Proximity ligation assay (PLA) showed that the elevated APAF1/Cytochrome C interaction (apoptosis), as shown by red fluorescent molecules, under H/R condition was antagonized by Irisin in JEG-3 cells, similar to our observation in human 1st trimester explants. (B) This process coincided with the inhibition of Caspase 3 activity as confirmed by specific fluorogenic substrates Ac-DEVD-AMC (n = 3). (Significant changes in Caspase 3 activity were measured by a one-way analysis of variance and subsequent Tukey’s post hoc test to analyze differences between cohorts; * p < 0.05. Scale bar = 100 μM. Blue fluorescent staining is for Dapi indicating the cell nucleus; Red fluorescent staining indicates APAF1 and Cytochrome C interaction. Bar plots are presented as mean ± SEM. N/T, non-treatment; PLA, Proximity Ligation Assay).