Figure 2

CGGBP1 depletion in U-2987 MG cells is associated with increased expression of CDKN1A and GAS1. A: A strong decrease in CGGBP1 mRNA expression shows the efficiency of siRNA transfections. Using GAPDH as control, increase in the mRNA levels of CDKN1A and GAS1 was seen. RNA samples from duplicate transfections were pooled and analyzed in triplicates and relative change in expression calculated by delta delta Ct method. All changes in expression are significant between control siRNA and CGGBP1 siRNA treatments (p < 0.05). Values for control siRNA in each case is normalized to 1 and not depicted. Plotted values are average ± S.D. and significance was calculated using heteroscedastic two-tailed T-test. B: Western blot analyses showed that CDKN1A and GAS1 expression were increased at protein level also. The mRNA levels of TP53 and ARF were also increased, but their protein levels were not increased as detected by western blots (data not shown). ACTB (top panel), CGGBP1 and GAS1 were run on the same blot. ACTB (lowest panel) and CDKN1A were run from same samples on a different blot.