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Fig. 2 | Molecular Cancer

Fig. 2

From: DDX3X loss is an adverse prognostic marker in diffuse large B-cell lymphoma and is associated with chemoresistance in aggressive non-Hodgkin lymphoma subtypes

Fig. 2

RNA-seq and protein network analysis of DDX3X mutant/depleted NHL cell subtypes and the molecular effect of DDX3X mutation/loss on DLBCL, NKTCL and CTCL cell lines. A Volcano plots showing DEGs in DDX3X-R475C mutant U2392, and DDX3X-depleted HuT78 and SNK1 cells (false discovery rate < 0.05 and p < 0.05) as analysed by RNA-seq (n = 3 each). Gene Ontology functional mapping of DEGs are shown in the corresponding insets. B Molecular interaction networks of DEGs that were identified by RNA-seq analysis of the above DDX3X-mutant/depleted cell lines, as predicted by the IPA®. Symbols representing protein types and their predicted relationships are provided in the legends. Solid and dashed lines represent direct and indirect interactions, respectively. C The mRNA levels of cyclin-D1 (CCND1) in U2392 cells expressing mutant DDX3X-R475C (left panel) or transfected with DDX3X shRNA (right panel) were quantified by RT-qPCR (WT, wild-type; CTL, control). D Cyclin-D1 expression levels in U2392 cells expressing mutant DDX3X-R475C (left panel) or transfected with DDX3X shRNA (right panel) were quantified by Western immunoblotting. Relative densitometry graphs quantifying cyclin-D1 expression are presented. E Control or DDX3X-mutant/depleted cell lines of DLBCL (U2392), NKTCL (SNK1, SNK6, NKYS) and CTCL (HuT78, MJ, MyLa) were analysed by Western immunoblotting to determine phosphorylation levels of STAT3 and p42/44. Relative densitometry graphs quantifying the expression levels of indicated proteins are presented. (F) U2392 cells with wild-type DDX3X or CRISPR knock-in mutant DDX3X-R475C were subcutaneously injected into NOD.Cg-PrkdcscidIl2rgtm1Wjll mice. Cells were left to grow for 3 weeks to develop into tumors. Xenografted mice were then injected (intra-tumoral) with vehicle or WP1066 (40 mg/kg) every 3 days for up to Day-58. Tumor volume (mm2) on each mice was recorded twice/week by caliper measurements and calculated by the modified ellipsoidal formula. n = 5 under each group. Data represent at least three experiments and values in graphs are mean ± SEM. ***, p < 0.001; **, p < 0.01; *, p < 0.05; ns, non-significant

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