Skip to main content
. 2018 Oct 22;128(12):5294–5306. doi: 10.1172/JCI98287

Figure 10. The effect of Ubqln1 overexpression on the degradation of a UPS surrogate substrate (GFPu) and autophagic flux in cultured cardiomyocytes.

Figure 10

(A–D) Cultured NRVMs were infected with Ad-Ubqln1 or Ad-Ubqln1-ΔUBL (or Ad–β-Gal as control) to overexpress Flag-tagged full-length or UBL domain–deleted Ubqln1, respectively. Representative images of Western blot analyses for the indicated proteins (A and C) and densitometry data pooled from 4 repeats (B and D). Ad-GFPu infection to express GFPu, a surrogate substrate for the UPS, was initiated at 24 hours after Ad-Ubqln1 infection; 48 hours later, total cell lysates were collected for Western blot analyses. (E and F) Autophagic flux assay. At 46 hours after Ad-Ubqln1 or Ad–β-Gal infection, bafilomycin A1 (BFA) or vehicle control (DMSO) was administered to inhibit lysosomes. Two hours later, cells were collected for Western blot analyses for the indicated proteins. β-Tubulin was probed for loading control. No Adeno, cells not subject to adenoviral infection. *P < 0.005; **P < 0.001, 2-tailed t test with Welch’s correction. Each dot represents a biological repeat.