A Cohesin-Independent Role for NIPBL at Promoters Provides Insights in CdLS
Figure 4
NIPBL is important to maintain gene activity.
Transcript levels of genes with NIPBL-bound promoters and no cohesin sites close to the gene (GLCCI1, BBX, TSPAN31, ARTS-1 and ZNF695) and the cohesin-regulated MYC gene were analyzed by RT-PCR/qPCR after RNAi depletion of NIPBL, MAU2 or SMC3 in HB2 cells. The cells were synchronized in G2 phase and the transcript levels are normalized against the housekeeping gene NAD. Transcripts of NIPBL, MAU2 and SMC3 were also analyzed to exclude that NIPBL affects transcription of MAU2 and SMC3 and vice versa. All three genes serve also as negative control genes without NIPBL binding site at the promoter, although MAU2 and SMC3 have intronic cohesin binding sites. P-values were determined using Students test using between 3 and 9 independent biological replicates. The p-value and number of replicates is indicated for each graph. Values that are significantly different (P-value<0.05) from control RNAi are highlighted in red. (error bars ± s.d.).