Figure 5. CaMKIV regulates a shift towards increased fission and reduced fusion during endotoxemia and CLP in vivo.

A-D, C57Bl/6J wild-type and CaMKIV^−/− (KO) mice were subjected to endotoxemia (5mg/kg) or cecal ligation and puncture; at the timepoints detailed, the mice were euthanized, and the kidneys harvested. A, total kidney protein lysate from 6 or 12 hours of CLP was analyzed by immunoblot. Densitometry of p-Ser⁶¹⁶-DRP1, p-Ser⁶³⁷-DRP1, OPA1, and Mfn1: solid bars = C57Bl/6J, open bars=CaMKIV^−/−. (representative blot of n=3 independent experiments; n= 4 (sham) 9–10 (CLP) mice total/group for both experiments combined). B, total mitochondrial protein lysate from 3 hours of endotoxemia was analyzed by immunoblot (representative blot of n=2 independent experiments. Densitometry of OPA1 and p-Ser⁶¹⁶-DRP1: solid bars = C57Bl/6J, open bars=CaMKIV^−/−. (n= 3–4 (control) 7–8 (endotoxemia) mice total/group for both experiments combined). C, 1μg DRP1 (molecular weight of recombinant protein, 104KD) was incubated in the presence or absence of CaMKIV (25 ng, molecular weight of recombinant protein, 70KD) and GSK3β (25 ng, molecular weight of recombinant protein, 38KD) for 10 min at 30°C with the following additions: 10mM MgCl₂, 0.2 mM ATP, 1 mM CaCl₂ and 1uM CaM. Reactions were terminated by boiling in SDS-2-ME dissociation solution and analyzed by immunoblot. (n=2 independent experiments). Data analyzed by Wilcoxon rank sum test.