Research Paper Volume 15, Issue 11 pp 4685—4698

Figure 2. CMA differs by sex, but not by age in UM-HET3 mouse liver lysosomes. (A) Representative western blots and quantifications are shown for the indicated proteins in 4 μg of the light (CMA+) lysosome fraction from the livers of ad libitum fed male and female mice of ages 4 and 24 months. (B) Representative western blots and quantifications are shown for the indicated proteins in 4 μg of the light (CMA+) lysosome fraction from the livers of male and female mice of ages 4 and 24 months that were fasted for 18 hours before euthanasia. (C) Representative western blots and quantifications are shown for a substrate binding and uptake assay using the light (CMA+) lysosome fraction from male and female mice of ages 4 and 24 months that were fasted for 18 hours before euthanasia. The right panel shows the fraction of broken lysosomes. In each case, fewer than 10% of lysosomes were broken. n = 3 for each group in every experiment. Statistical analysis was performed in GraphPad Prism 9. Lines are drawn at each mean, with error bars showing S.E.M. p-values derived from 2-way ANOVAs are shown beneath each graph. “Estimation plots” are shown to the right graphs for LAMP2A and HSPA8 (the two proteins most important for CMA activity) Error bars on estimation plots show the 95% C.I. for the difference between the means of the indicated groups. p values displayed directly on the graphs are derived from unpaired t tests.