Figure 5

Myocardial hypothermia increases mitophagy. Female farm pigs (n = 3 per group) were euthanized one week post MI. WB analysis were performed on the mitochondrial fraction of myocardial tissue. All proteins were normalized to ponceau. (A) WB analysis with protein quantification (B–G) of parkin, Optineurin, p62, NDP52, LC3-II and phospho-Ubiquitin (Ser65) respectively in the mitochondrial fraction. Our hypothesis of mitophagy is supported by a strong increase of the short form of OPA-1 and MFF, as well as a significant increase of DRP-1 in the whole lysate (H–J) The data represents the mean ± SEM with *p < 0.05, comparing cold (black) vs. warm (white). (K) Representative EM images (3 h post MI) of) mitophagy (x100000) and damaged mitochondria (thin black arrow) with mitophagy (thick black arrow) (x19000) under cold condition. We normalized each protein to the entire corresponding lane on the ponceau-stained membrane, representing the total protein amount. Due to space limitation, the blots are cropped from various gels, please see the supplemental data for full-length gels/blots.