fig4

Figure 4. GSK-3β^S9A knock-in (βKI) attenuates aging-induced decreases in autophagy. (A) Immunoblots of LC3, p62, and α-tubulin in left ventricular homogenates of WT and homozygous βKI mice at 6 and 24 months (M). (B) Relative band density of LC3-II/tubulin. (C) Relative band density of p62/tubulin. In (B, C), WT6, WT 6M; βKI6, βKI 6M; WT24, WT 24M; βKI6, βKI 6M. The relative band density of WT6 was designated as 1. *P < 0.05 vs. WT6; ^#P < 0.05 vs. WT of the same age. n = 6 in each group. (D) Immunoblots of LC3, p62, and tubulin in cardiomyocytes transduced with LacZ or GSK-3β, with or without chloroquine treatment (CQ, 20 µM) for 4 h. The results of two experiments (out of four) are shown. (E) Relative band density of LC3-II/tubulin. (F) Relative band density of p62. In (E, F), the relative band density in LacZ-transduced cells treated with vehicle was designated as 1. ^#P < 0.05 vs. LacZ of the same treatment; *P < 0.05 vs. vehicle control (Ctrl) with the same virus infection. n = 4 in each group. (G) GFP-LC3 dots in cardiomyocytes transduced with GFP-LC3 and LacZ or GSK-3β, with or without chloroquine treatment (CQ, 20 µM) for 4 h. The number of GFP-LC3 dots is quantified in (H). In each experimental repeat, 20 cardiomyocytes were counted. ^#P < 0.05 vs. LacZ of the same treatment; *P < 0.05 vs. vehicle control (Ctrl) with the same virus infection. WT: Wild type.