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. 2016 Apr 19;12(4):e1005895. doi: 10.1371/journal.pgen.1005895

Fig 4. SRSF1 is required for tumorigenecity of SCLC.

Fig 4

(a) and (b): DMS114 cells were transfected with non-targeting control or SRSF1-directed siRNAs for 48 hrs, then treated with cisplatin (2.5ug/ml) or topotecan (2.5ug/ml) for 24 hrs. Cell growth (a) and Caspase-3/7 activities (b) were assessed and normalized against non-targeting ctrl siRNA-transfected cells as 100% control. (c): DMS114 cells were transfected with non-targeting and SRSF1 siRNAs for 48 hrs and then seeded in sphere forming media and allowed to grow for 4 days. Phase-contrast images of the sphere formation under each condition were captured and viable cell mass quantitated by CTG assay. (d): Reconstitution of SRSF1 expression using a siRNA-resistant Flag-tagged SRSF1 expression construct was carried out in SRSF1 siRNA transfected cells. Impact on sphere growth rate was assessed by CTG assay, and successful SRSF1 protein re-expression was confirmed using either anti-SRSF1 antibody or anti-Flag antibody. (e) DMS114 cells transfected with non-targeting control siRNA or SRSF1 siRNA were implanted into immunocompromised mice and tumor formation rates were monitored and measured as described in Materials and Methods.