Fig. 5 | Nature Communications

Fig. 5

From: Hsp70 and Hsp40 inhibit an inter-domain interaction necessary for transcriptional activity in the androgen receptor

Fig. 5

Hsp70-targeted small molecules increase ubiquitination and degradation of polyQ-AR. a PC12 cells were transiently transfected to express HA-ubiquitin and CHIP, and AR112Q expression was induced for 48 h. Cells were treated with the Hsp70 modulator, JG-98 (0.5 μM) for the last 24 h and with 10 μM MG132 for the last 16 h. AR112Q was immuno-precipitated from lysates and then probed for ubiquitin (HA). Left: input. Middle: pull down. Right: quantification from three independent experiments. Data are mean ± S.E.M. from three independent experiments. *p < 0.05 by two-tailed t-test. b JG-98 promotes AR112Q degradation by the proteasome. PC12 cells were induced to express AR112Q for 48 h in the presence of R1881 (10 nM) and JG-98 (0.5 or 1.0 μM). Indicated samples were treated with lactacystine (10 μM) for the final 16 h. AR was detected by western blot. GAPDH controls for loading. c JG-98 does not induce a stress response. PC12 cells were induced to express AR112Q in the presence of R1881 (10 nM) for 48 h in the presence of 17-AAG (5 μM), JG-98 (0.5 μM) or vehicle control. Cell lysates were probed for Hsp25, Hsp40, Hsp70, Hsp90, Akt, and ERK1/2. GAPDH controls for loading. d PC12 cells were induced to express AR112Q in the presence of R1881 for 48 h, and treated with 17-AAG (1 µM) and/or JG-98 (1 µM), as indicated. Lysates were analyzed for AR by filter trap assay. Data are mean ± S.E.M. from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA. Source data for the quantifications shown in a, d are available as Source Data file

Back to article page