Figure 3.

PARP1 and PARylation affects total mRNA levels differentially. Total mRNA levels using exonic primers (green arrows) were measured at each stage of the experiment. WT for and PARP1 KD cells were treated with PJ34 overnight, to inhibit PARylation. Control cells are cells in each condition (WT or KD) that was not treatment with PJ34 at each stage. At the beginning (Stage 1 or S1), mRNA levels were measured from cells. Next, cells were incubated with 30 μg/ml DRB for 3 hrs (Stage 2 or S2); then DRB containing medium was removed, fresh medium was added and transcription was allowed to resume (Stage 3 or S3). At the stage of recovery (S3), levels of total RNA as measured by RT-PCR using exonic primers (green arrows) were determined for AKAP200 (A) and CAPER (B) and PARP1 KD cells for AKAP200 (C) and CAPER (D). mRNA levels were normalized to the values prior to PJ34 treatment sample, which was set to 1. Results are shown as mean ± SEM from three independent experiments. All results were measured in relation to the non-treated cells in each condition (WT or KD) and results were significant as analyzed using student T-test p < 0.05.