Reduction of CTSL mRNA in acetaldehyde treated HepG2 cells.

<p>Total cellular RNA isolated from acetaldehyde treated or untreated cells was reverse transcribed and subjected to real time PCR using sybergreen and gene specific primers for (A) CTSL (B) cathepsin B or (C) cystatin C. Similarly real time PCR for 18 S rRNA was performed and used as an internal control. Cycle threshold (Ct) values were calculated for each PCR and relative fold change was calculated using 2−^{ΔΔ Ct} method. Each set of observation was compared to the other set using a paired two-tailed t-test, assuming unequal variances among the sample means. A p value of ≤0.05 was considered to be statistically significant. Values are mean ± SD from three independent experiments performed in triplicates. Values significantly different from control has been marked by *. (D) Analysis of PCR products. After the completion of real time PCR amplified products were resolved on agarose gel to confirm the amplification of a single product. Representative gel for each target is given in the figure.