Fig. 1

HuD is required for cancer cell survival. A HuD protein expression analysis in peripheral nerve tissue (normal) vs. neuroblastoma (cancer); n is the total number of samples in the patient tissue array (the array represents 27 cases of neuroblastoma, plus 5 cases of normal peripheral nerve tissue) (see Methods under “Immunocytochemistry (ICC), immunohistochemistry (IHC-F), and tissue array staining.” The array content is listed in Table S9 of the supplementary methods). Scale bar corresponds to 200 μm. Relative protein quantifications are shown (right). B HuD expression across different cancer cell lines obtained from CCLE; data are presented as mean ± SD. C Expression quantification for HuD in human neurons and neuroblastoma by RT-qPCR. D and E HuD shRNA treatment reduced HuD massage and protein expression in IMR-32 and SK-N-SH cells. Full-length blots are presented in Supplementary Fig. S9. F and G Validation of HuD shRNA and shRNA inactive HuD mutant constructs with viability and Western blotting in IMR-32 cells. Full-length blots are presented in Supplementary Fig. S9. H Viability assay (control or silenced HuD and/or overexpressed HuB and/or overexpressed HuC) in IMR-32 cells. I Validation of viability loss in shHuD inducible IMR-32 and SK-N-SH cells. J Viability assay comparison between HuD over-expressers - IMR-32, NCI-H146, NCI-H69, NCI-H889, NCI-H209, and low expresser - SK-N-MC (with control or silenced HuD). Data are presented as mean ± SEM; t-test: *p < 0.05, **p < 0.01, ***p < 0.001