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. Author manuscript; available in PMC: 2012 Jun 15.

Published in final edited form as: Toxicol Appl Pharmacol. 2011 Apr 3;253(3):235–243. doi: 10.1016/j.taap.2011.03.024

Fig. 1 — Negative control of p63 expression in keratinocytes by nickel treatment. (A) and (B), Down-modulation of p63 mRNA expression by nickel. Both HaCat and HPK cells were treated with different doses of nickel for 24 hr (A), and treated with 1.0 mM for different time (B). ΔNp63 mRNA levels were quantified by real time RT-PCR. Values are expressed as relative arbitrary units, after internal normalization for GAPDH mRNA expression. (C) Down-modulation of p63 protein expression by nickel. Both HaCat and HPK cells were treated with nickel for different times and doses. The cells then were analyzed for ΔNp63 protein level by immunoblotting with the corresponding antibodies. Immunoblotting for β-actin was used for equal loading control. (D) Up-regulation of both p53 and p73 by nickel treatment. HaCat cells were treated with nickel for 24 hr. The cells were harvested, p53^ser15, phospho-p73 and β-actin expression were analyzed by immunoblotting. (E) Down-modulation of p63 activity in keratinocytes in response to nickel treatment. Both the HaCat and HPK cells were transfected with 0.5 μg of Np63 luciferase reporter DNA followed by nickel treatment. The cells were harvested for the measurement of luciferase activity after 24 hr. Values are expressed as relative units after internal normalization for protein concentration. *, p<0.05 compared to control without nickel treatment (one-way ANOVA test).

Fig. 1 — Negative control of p63 expression in keratinocytes by nickel treatment. (A) and (B), Down-modulation of p63 mRNA expression by nickel. Both HaCat and HPK cells were treated with different doses of nickel for 24 hr (A), and treated with 1.0 mM for different time (B). ΔNp63 mRNA levels were quantified by real time RT-PCR. Values are expressed as relative arbitrary units, after internal normalization for GAPDH mRNA expression. (C) Down-modulation of p63 protein expression by nickel. Both HaCat and HPK cells were treated with nickel for different times and doses. The cells then were analyzed for ΔNp63 protein level by immunoblotting with the corresponding antibodies. Immunoblotting for β-actin was used for equal loading control. (D) Up-regulation of both p53 and p73 by nickel treatment. HaCat cells were treated with nickel for 24 hr. The cells were harvested, p53^ser15, phospho-p73 and β-actin expression were analyzed by immunoblotting. (E) Down-modulation of p63 activity in keratinocytes in response to nickel treatment. Both the HaCat and HPK cells were transfected with 0.5 μg of Np63 luciferase reporter DNA followed by nickel treatment. The cells were harvested for the measurement of luciferase activity after 24 hr. Values are expressed as relative units after internal normalization for protein concentration. *, p<0.05 compared to control without nickel treatment (one-way ANOVA test).