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. 2013 May 27;169(4):794–807. doi: 10.1111/j.1476-5381.2012.02208.x

Figure 7.

Figure 7

Role of MAPK in URB597 control of TH promoter activity. The influence of MAPK signalling was investigated in N1E115 cells transfected with pTH250-Luc. To examine the role of this signalling cascade, cells were treated for 1 h with MEK1/2 inhibitor U0126 (5 μM) before adding URB597 (0.1 μM) for an additional 5 h (A). Results are given as the percentages of relative luciferase activity (firefly luciferase relative to Renilla luciferase) relative to control values. Data shown are means with SEM values of three experiments performed in triplicate. One-way ANOVA indicated a general effect of the MEK1/2 inhibitor (***P = 0.0003, f = 36.33, residual d.f. = 8) with a positive interaction (*P = 0.0139, f = 9.831). ##P < 0.01 as indicated for treated cells and determined by Bonferroni post-test. (B) ERK1/2 activation was determined using immunoblot detection of phosphorylated ERK1/2 (P-ERK1/2 – 42 and 44 kDA). Cells were treated with URB597 (1 μM, 15 min) in the presence or the absence of SR 141716A (1 μM). Phosphorylated ERK1/2 values were normalized to total ERK1/2 (Tot ERK1/2). Results are expressed as percentages of relative density. A representative immunoblot is shown in (C). Data shown are means with SEM values of three experiments. One-way ANOVA indicated a general effect of URB597 (**P < 0.0016, f = 21.80, residual d.f. = 8). Neither significant effect for SR 141716A nor an interaction were observed, indicating the absence of antagonism by SR 141716A.