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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 3. TNF-α treatment significantly decreases Prepro-Hypocretin expression in B35 cells transfected with MSCV-Prepro-Hypocretin vector.

Figure 3

B35 cells overexpressing Prepro-Hypocretin (mediated by MSCV-Prepro-Hypocretin vector) or overexpressing GFP control protein (mediated by MSCV-GFP) were cultured in the presence and absence of TNF-α cytokine (2 ng/ml) for 24 hours at 37°C. Panels A and E: Cells were lysed, and equivalent amount of whole cell detergent lysates were Western blotted with the indicated antibodies. FAK and G3PDH protein expression served as controls. Panel B: Densitometry of Prepro-Hypocretin expression as shown in Panel A (normalized to G3PDH and relative to the controls: the cells treated with vehicle only) as described in the Materials and Methods 2.3. Panels C and D: Total RNA was extracted from the above cells to analyze the mRNA level of Prepro-Hypocretin (in Panel C) and FAK (in Panel D as a control) by quantitative real time RT-PCR. Data was normalized to beta-actin and relative to the control (cells treated with vehicle only). For Panels B and C, open bar denotes the results from control cells (vehicle only), and Black bar denotes the results from TNF-α-treated cells. Data are presented as mean ± S.D. (n = 4). * represents p < 0.01 for TNF-α-treated cells compared to vehicle-treated control group.