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. 2010 Feb 22;285(17):13254–13263. doi: 10.1074/jbc.M110.103051

FIGURE 2.

FIGURE 2.

Effect of overexpression of HSP70 on IBMX-stimulated activity and expression of tyrosinase. HSP70-overexpressing B16 cells (HSP70 +) and mock transfectant control cells (HSP70 −) were incubated for 48 h with or without 100 μm IBMX (A–C). Tyrosinase activity was determined as described under “Experimental Procedures” and expressed relative to the control (A). Whole cell extracts were analyzed by immunoblotting as described in the legend of Fig. 1B. Total RNA was extracted and subjected to real-time RT-PCR using a specific primer for the tyrosinase gene. Values were normalized to gapdh gene expression and expressed relative to the control sample (C). HSP70-expressing B16 cells (HSP70 +) and mock transfectant control cells (HSP70 −) were transiently transfected with pRL-SV40 (internal control plasmid carrying the R. reniformis luciferase gene) and pGL4-tyrosinase-luc. After 24 h, the cells were incubated for 24 h with or without 100 μm IBMX. P. pyralis luciferase activity was measured and normalized for R. reniformis luciferase activity (D). Values are given as the mean ± S.D. (n = 3). **, p < 0.01.