Figure 2.
Pol II phosphorylation mediates cotranscriptional splicing of an RBM39 poison exon. (A) Western blot to detect changes in phosphorylated SR-rich proteins after treatment of NALM6 cells with increasing concentrations of EHT1610 for 8 hours. (B) Western blot analysis of extracts from NALM6 cells treated with DMSO, DRB (30 μM), and EHT1610 for 4 hours. IIo and IIa indicate hyperphosphorylated and hypophosphorylated CTD52, respectively. (C) Pie chart showing differential splicing events in NALM6 cells treated with DMSO when compared with those treated with THAL-SNS-032 (500 nM) for 8 hours. (D) The inclusion level in NALM6 cells treated with DMSO (n = 3) and THAL-SNS-032 (n = 3) for the identical alternative spliced event in RBM39 transcript shown in Figure 1F. ∗P < .05. (E) The NALM6 cells were treated with increasing doses of THAL-SNS-032 for 8 hours. PCR reactions were performed for the detection of RBM39 splicing events (upper), and western blot analysis of extracts were shown (lower). (F) NALM6 cells were treated with EHT1610 in a time-dependent manner. PCR reactions were performed for the detection of the RBM39 splicing event (upper), and western blot analysis of extracts were shown (lower). (G) NALM6 cells were treated with EHT1610 (4 μM) for 4 and 24 hours. PCR reactions were performed for detection of the RBM39 splicing event (upper), and western blot analysis of extracts are shown (lower). (H) NALM6 cells were treated with EHT1610 for 16 hours, followed by additional treatment of DRB for 4 hours. PCR reactions were performed for the detection of RBM39 splicing event (upper), and western blot analysis of extracts were shown (lower). (I) NALM6 cells (upper) or MUTZ-5 cells (lower) were treated with EHT1610 for 16 hours, followed by additional treatment with THAL-SNS-032 for 4 hours. PCR reactions were performed to detect the RBM39 splicing event. (J) NALM6 cells were treated with EHT1610, THAL-SNS-032 or a combination for 6 hours. PCR reactions were performed for the detection of an RBM39 splicing event (upper) and representative western blot analysis of 3 biological replicates of extracts are shown (lower).

Pol II phosphorylation mediates cotranscriptional splicing of an RBM39 poison exon. (A) Western blot to detect changes in phosphorylated SR-rich proteins after treatment of NALM6 cells with increasing concentrations of EHT1610 for 8 hours. (B) Western blot analysis of extracts from NALM6 cells treated with DMSO, DRB (30 μM), and EHT1610 for 4 hours. IIo and IIa indicate hyperphosphorylated and hypophosphorylated CTD52, respectively. (C) Pie chart showing differential splicing events in NALM6 cells treated with DMSO when compared with those treated with THAL-SNS-032 (500 nM) for 8 hours. (D) The inclusion level in NALM6 cells treated with DMSO (n = 3) and THAL-SNS-032 (n = 3) for the identical alternative spliced event in RBM39 transcript shown in Figure 1F. ∗P < .05. (E) The NALM6 cells were treated with increasing doses of THAL-SNS-032 for 8 hours. PCR reactions were performed for the detection of RBM39 splicing events (upper), and western blot analysis of extracts were shown (lower). (F) NALM6 cells were treated with EHT1610 in a time-dependent manner. PCR reactions were performed for the detection of the RBM39 splicing event (upper), and western blot analysis of extracts were shown (lower). (G) NALM6 cells were treated with EHT1610 (4 μM) for 4 and 24 hours. PCR reactions were performed for detection of the RBM39 splicing event (upper), and western blot analysis of extracts are shown (lower). (H) NALM6 cells were treated with EHT1610 for 16 hours, followed by additional treatment of DRB for 4 hours. PCR reactions were performed for the detection of RBM39 splicing event (upper), and western blot analysis of extracts were shown (lower). (I) NALM6 cells (upper) or MUTZ-5 cells (lower) were treated with EHT1610 for 16 hours, followed by additional treatment with THAL-SNS-032 for 4 hours. PCR reactions were performed to detect the RBM39 splicing event. (J) NALM6 cells were treated with EHT1610, THAL-SNS-032 or a combination for 6 hours. PCR reactions were performed for the detection of an RBM39 splicing event (upper) and representative western blot analysis of 3 biological replicates of extracts are shown (lower).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal