Figure 4.

p62 is the common downstream target of anesthetic drugs on tooth pulp cells. (a) Heatmap analysis of PCR array data showing changes in gene expression in tooth pulp cells after 16 h treatment with indicated 2 mM anesthetic drugs (for 0.5 mM dosage, please see Supplementary Figure 4). Colors represent fold changes in expression, as shown in the key. (b) Real-time RT-PCR analysis of a panel of autophagy-related genes in primary tooth pulp cells using specific primers, normalized to 36β4 housekeeping gene expression. Error bars represent standard deviation of the triplicate experiments. (c) Western blot analysis of p62 expression in drug-treated cells. β-actin was used as a loading control by stripping and reblotting the same blot. p62 signal quantification was calculated with normalization against β-actin signal (right panel). Similar results have been achieved with lidocaine both for western blot and gene profiling (data not shown). UBI, Ubistesin; UBI-F, Ubistesin forte; Scan, Scandonest; Sept, Septanest. *P<0.05; **P<0.01. (d) Real-time RT-PCR analysis of p62 gene in tooth pulp cells treated with the five anesthetics for different time periods, normalized to 36β4 housekeeping gene expression. Error bars represent standard deviation of triplicate experiments.