Suppression of βTRCP increases TRIB1 expression but does not affect its instability.

HeLaT1 cells incubated for 48 h with siRNA targeting CUL1, βTRCP or a non-target control were treated with vehicle (H20) or CHX for 5 h. Parallel lysates were then analyzed by western blot and qPCR. A, qRT-PCR of RNA isolated from cells treated with the indicated siRNAs for 48 h. Quantification of corresponding transcript is shown, normalized to the non-target oligonucleotide (NT). B, western blot of extracts from silenced cells. Data shown are representative of 3 biological replicates. C, βTRCP and CUL1 suppression increase TRIB1 transcript abundance. qRT-PCR of TRIB1 in cells suppressed for the indicated siRNAs. In A and C, data represent the average of 2 biological replicates ± S.D.