Fig. 6.

CAL inhibits rotenone-induced glutamate release to modulate mGluR5 activity. (A–D) Cells infected with LV-CAL or LV-NC were exposed to rotenone followed by DHPG treatment, and the alteration in protein levels of mGluR5 (A), p-JNK (B), TH (C), and cell viability (D) was evaluated. Protein levels shown in corresponding graphs were normalized to GAPDH or total JNK and represented as the fold difference of the control group. (E) Cells were transfected with CMV-hCAL-IRES-ZsGreen or vector CMV-IRES-ZsGreen followed by 0.2 μM rotenone treatment for 24 h and then the supernatant was assessed for glutamate concentration. (F, G) Cells transfected with shGFP or shCAL were pretreated with MPEP (100 μM, 30 min) followed by rotenone for 24 h, the supernatant was obtained for the glutamate assay (F), and the cell lysates were probed with anti-mGluR5 and anti-CAL antibodies by Western blotting (G). Protein levels were normalized to GAPDH and represented as the fold difference of the control group. LV-NC infected cells without rotenone treatment were used as controls (A–D). Cells transfected with vector without rotenone administration were used as controls (E–G). Data shown in all panels of this figure represent the mean ± SEM of 3 independent experiments. The statistical significance was determined by 1-way ANOVA followed by Dunnett’s test. **p < 0.01 and ***p < 0.001 versus control, ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 compared to the rotenone treatment group, ^&p < 0.05 and ^&&p < 0.01 compared to the rotenone-treated LV-CAL group or rotenone- and MPEP-treated shGFP group