Fig 2: DHX38 promotes splicing of the HBB-derived reporter.(A) A representative gel of HBB-derived reporter splicing after DHX38 knockdown and expression of siRNA-resistant DHX38WT-FLAG and DHX38G332D-FLAG. (B) A scheme of HBB-derived reporter and M1 and M2 mutants that block usage of normal splice site and promote usage of upstream 5’ cryptic splice site. (C) Quantification of three independent experiments with the mean and SEM. Statistical significance was assessed by two tail unpaired t-test (* indicates p≤0.05, ** p≤0.01).

Fig 3: DHX38 associates with multiple spliceosomal components.DHX38WT-FLAG and DHX38G332D-FLAG were transiently expressed in HEK293 cells, immunoprecipitated and copurification of selected spliceosomal proteins monitored. Immunoprecipitation was performed in normal NET2 buffer containing 150 mM NaCl (A) or in stringent NET2 buffer containing 300 mM NaCl (B).

Fig 4: DHX38 downregulation changes primarily gene expression.Summary of three independent RNA-seq analysis of HEK293 cells treated either with anti-DHX38 siRNA or negative control siRNA. Analysis of (A) splicing efficiency, (B) alternative splicing and (C) gene expression. Expression of selected genes that were downregulated (D) or upregulated (E) after DHX38 downregulation was analyzed by RT-qPCR. RNA was isolated from control cells or cells treated with cycloheximide (CHX) to reveal potential targets of non-sense mediated decay. Average of three independent experiment with SEM is shown. Expression value determined by RNA-seq analysis is shown in the same graph for comparison (RNA seq).