FIG. 9.

Signaling mechanisms involved in HDRP-mediated neuroprotection. (A) Effect of HDRP expression on various signaling proteins. Neuronal cultures infected with the Ad-HDRP or Ad-GFP expression vector were treated with HK or LK medium for 3 h, and protein extracts were prepared and subjected to Western blot analysis using different antibodies sequentially on the same membrane. Equal protein loading was confirmed by probing the membrane with α-tubulin antibody. (B) c-Jun expression in HDAC9^−/− neuronal cultures during forced expression of HDRP. Neuronal cultures were prepared from HDAC9^−/− mice and infected with Ad-HDRP or Ad-GFP. Cells were then treated with HK or LK medium for 3 h. (Top panel) RNAs were extracted from treated neurons, and RT-PCR was performed using primers specific to the c-Jun transcript. Equal sample loading was ensured by examining GAPDH levels. (Bottom panel) Cellular protein was extracted from treated neurons, and Western blotting was performed. The membrane was then probed with antibodies against c-Jun and HDAC9. Equal protein loading was confirmed by probing the membrane with α-tubulin antibody.