Northern blot hybridization analysis of differential expression of α1-ACT, HEF-1α, COX-II, and OAz genes in PCI-06A-Raf(AS) versus PCI-06A-Raf(K) transfectants
Top panels: Northern blots were sequentially hybridized first with a radiolabeled partial human cDNA probe (KAS-101, α1-ACT; KAS-102, HEF-1α; KAS-103, COX-II; or KAS-104, OAz), followed by the radiolabeled GAPDH cDNA probe. The sizes of various transcripts are shown in parentheses. Lower panel: Computerized densitometric scanning analysis of the relative increase in gene expression. Relative fold increase in the steady-state mRNA level was calculated by first normalizing against the GAPDH signal, followed by comparison with the expression in PCI-06A-Raf(K) transfectants (α1-ACT and COX-II) or PCI-06A-Raf (AS) transfectants (HEF-1α and OAz), as explained in Materials and Methods. The data were analyzed using the paired t-est (Excel version 5.0), and the p values for the statistical significance of the difference between the AS and the K groups were <0.05 (α1-ACT, HEF-1α, COX-II). AS, PCI-06A-Raf(AS); K, PCI-06A-Raf(K).