IL-32γ induces sustained steady state levels of MCL-1 protein.

<p>Human neutrophils were incubated for 8, 20 and 30 hrs with IL-32γ (300 ng.ml⁻¹) or its vehicle (absence of IL-32γ). Cell lysates were subjected to SDS-PAGE and transferred to PVDF membranes. Western blotting was then performed with anti-MCL-1 or anti-β-actin (control of protein loading) antibodies, and proteins were revealed with Western Lightning Plus ECL. Representative of 4 different donors (<b>A</b>). Densitometric analyses of MCL-1 and actin expression were performed and ratios between MCL-1 and actin were calculated to normalize the data. These normalized values were then used to calculate the percentages of MCL-1 variations in neutrophils incubated with vehicle at 20 and 30 hrs of incubation or with IL-32γ at 8, 20 and 30 hrs of incubation, considering that the normalized value of MCL-1 in neutrophils incubated with vehicle at 8 hrs represents 100% MCL-1 (neutrophils at 8 hrs of incubation with vehicle were 96% non-apoptotic) (<b>B</b>). Results are means ± s.e.m. of 4 different donors. Statistics: paired Student <i>t</i> test, * p<0.05.