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. Author manuscript; available in PMC: 2012 Apr 1.
Published in final edited form as: Biochim Biophys Acta. 2010 Nov 18;1812(4):565–571. doi: 10.1016/j.bbadis.2010.11.003

Figure 1. TNF-α treatment significantly decreases endogenous Prepro-Hypocretin mRNA and protein levels, as well as endogenous HcrtR2 protein level in rat B35 cells.

Figure 1

Figure 1

Rat B35 neuroblastoma cells were cultured in the presence and absence of TNF-α cytokine (2 ng/ml) for 24 hours at 37°C. Panel A: Low levels of endogenous Prepro-Hypocretin and HcrtR2 were detected by RT-PCR as described in the Materials and Methods in B35 cells without TNF-α treatment. Reactions without reverse transcriptase (RTase) were PCR controls. Panels B, D, and F: Total RNA in B35 cells treated with or without TNF-α was extracted to analyze the Prepro-Hypocretin mRNA level in Panel B, the HcrtR2 mRNA level in Panel D, and the control FAK mRNA level in Panel F, by quantitative real time RT-PCR (normalized to beta-actin and relative to the controls: the cells treated with vehicle only). Panels C and E: The above B35 cells were lysed, and equivalent amount of whole cell detergent lysates were Western blotted to analyze the protein level of Prepro-Hypocretin and HcrtR2 with the indicated antibodies. FAK and G3PDH expression served as controls.