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. 2020 Sep 2;8:859. doi: 10.3389/fcell.2020.00859

FIGURE 4.

FIGURE 4

Effects of proteasome dysfunction on the proteolytic system. (A) Immunoblotting analysis of ubiquitinated proteins in the tibialis anterior muscles of Con and mKO mice after 3 weeks of doxycycline (DOX) treatment. GAPDH served as a loading control. (B) Immunostaining of cryosections of the tibialis anterior muscles of Con and mKO mice for laminin α2 (red), LC3 (green), and DAPI (blue) after 3 weeks of DOX treatment. Scale bar = 50 μm. (C) Immunostaining of cryosections of the tibialis anterior muscles of Con and mKO mice for laminin α2 (red), p62 (green), and DAPI (blue) after 3 weeks of DOX treatment. Scale bar = 50 μm. (D) Immunoblotting analysis of autophagy-related proteins in the tibialis anterior muscles of Con and mKO mice after 3 weeks of DOX treatment. (E) Quantification of LC3II and p62 proteins and the LC3II/LC3I ratio. Data represent the means ± SEM (t-test: *p < 0.05; n = 3 per group). (F) Graphical summary of the study. Con indicates Rpt3f/f mice, and mKO indicates muscle-specific Rpt3-knockout mice (ACTA1-rtTA;tetO-Cre;Rpt3f/f).