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. 2016 Oct 6;6:34784. doi: 10.1038/srep34784

Figure 5. MB down regulates MARK4 protein level through ubiquitin-proteasome degradation in 293T cells.

Figure 5

(a) Representative immunoblot of lysates from 293T cells transfected with HA-MARK4-KD or WT. Cells transfected with HA-MARK4-WT were treated with AC, MB and OS as described in Fig. 2a. MARK4 protein level was evaluated using an anti-HA antibody with β-actin as the loading control. Lower panel shows the quantification of HA-MARK4-WT normalized to β-actin presented as the percent of the level found in cells transfected with MARK4-WT but without drug treatment. Results are mean ± SD (n = 3). *0.01 < P < 0.05; **0.001 < P < 0.01; ***P < 0.001. Student’s t test is conducted between corresponding values in AC and OS or MB and OS group. (b) Representative immunoblot of immunoprecipitated HA-MARK4 probed by ubiquitin antibody (WB: Ub) and HA antibody (WB: HA). HA-MARK4 was immunoprecipitated from lysates of 293T cells transfected with HA-MARK4 followed by incubation with increasing concentrations of MB for 3 h. (c) Representative immunoblot of HA-MARK4 from lysates of 293T using an anti-HA antibody with β-actin as the loading control. Cells transfected with HA-MARK4 were treated with increasing concentrations of MB for 3 h in the presence of absence of 500 nM of proteasome inhibitor (R)-MG132. All blots were cropped. Full blots are shown in Supplementary Fig. S3.