Isoginkgetin and Madrasin are poor splicing inhibitors
Fig 5
Isoginkgetin treatment affects pol I and pol II transcription.
(A) qRT-PCR with primers amplifying different protein-coding genes. HeLa cells were treated with DMSO or 30 μM Isoginkgetin for 1h to 6h. cDNA was generated with oligo(dT). Values are normalised to the protein-coding gene GAPDH and shown as relative to DMSO, mean ± SEM, n = 4 biological replicates. Statistical test: two-tailed unpaired t-test. P-value: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001. (B) Total pol II ChIP-qPCR on the intron-containing gene KPNB1 in HeLa cells treated with DMSO or 30 μM Isoginkgetin for 1h to 6h. Mean ± SEM, n = 3 biological replicates. Statistical test: two-tailed unpaired t-test. P-value: * < 0.05, ** < 0.01. (C) Total pol II ChIP-qPCR on the intronless gene JUN in HeLa cells treated with DMSO or 30 μM Isoginkgetin for 1h to 6h. Mean ± SEM, n = 3 biological replicates. Statistical test: two-tailed unpaired t-test. P-value: * < 0.05. (D) Total pol II ChIP-qPCR of the histone gene H1-2 in HeLa cells treated with DMSO or 30 μM Isoginkgetin for 1h to 6h. Mean ± SEM, n = 3 biological replicates. Statistical test: two-tailed unpaired t-test. P-value: * < 0.05. (E) Western blots of total pol II, Ser2-P, Ser5-P, and histone H3 on the chromatin fraction of HeLa cells treated with DMSO or 30 μM Isoginkgetin for 1h, 2h, or 4h. Quantifications of the western blots are shown below each panel. Each quantification has been normalised to histone H3 signal and then to the DMSO condition. (F) ChIP-qPCR of total pol II, Ser2-P, and Ser5-P across the intron-containing gene KPNB1 in HeLa cells treated with DMSO or 30 μM Isoginkgetin for 1h, 1h30, or 2h. Mean ± SEM, n = 3 biological replicates. Statistical test: two-tailed unpaired t-test. P-value: * < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001.