Fig. 8: RBM39 controls its homoeostasis through non-canonical 3′-splice site selection.
From: Molecular basis of RNA-binding and autoregulation by the cancer-associated splicing factor RBM39
a Annotated sequence of the RBM39 exon 2b and its flanking regions. Below, the predicted inhibitory secondary structure SLi is displayed as well as the ΔBS2 and the optSLi stem loops. b Schematic representations of the different minigene constructs (WT, ΔBS2, HBB 3′ss, mutBS1 and optSLi). c Western blot analysis of the RBM39 knock down. GAPDH served as a loading control (n = 3). d Agarose gel showing the results of the RT-PCR using the different minigene constructs (WT, ΔBS2, HBB 3′ss, mutBS1 and optSLi) in control and RBM39 KDs. e Barplot showing the percentage of poison exon inclusion as determined by RT-PCR for the different minigene variants upon Ctlr and RBM39 KD. Average values and standard deviations of three biological replicates (n = 3) are shown. P values were computed using two-sided t test. f Plot showing the amide chemical shift perturbations observed upon titration of RBM39 RRM2 with BS1 or mutBS1. The protein was concentrated to 200 μM and the RNA stock was at 2 mM.
