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. 2017 Feb 15;7:42504. doi: 10.1038/srep42504

Figure 5. The antitumor activity of JA is mediated by SF3B1 and SF3B3.

Figure 5

(A–D) A stable pool of isogenic cells depleted for SF3B1 or SF3B3 was generated in MCF-7 and MDA-MB-468 cells using two independent lentiviral shRNA constructs targeting the endogenous proteins, followed by brief puromycin selection. Levels of knock-down were evaluated by immunoblotting. SF3B1 or SF3B3 depleted cells were treated with 1 μM of JA. Apoptosis was evaluated at 72 h after treatment. Cropped blot is shown from one representative experiment. Full-length gels are included in the Supplementary information file. Bars represent the means ± s.d. of 3 independent experiments. Asterisks (*) indicate statistical significance compared with control cells without JA treatment (P < 0.01, Student’s t-test). Vec, vector control cells. NS, non-targeting shRNA. (E and F) Simultaneous knock-down of SF3B1 and SF3B3 completely abrogated the apoptotic effects of JA. MCF-7 cells were transfected with SF3B1si-1 and/or SF3B3si-1 for 48 h followed by JA treatment for 24 h. Cropped blot is shown from one representative experiment. Full-length gels are included in the Supplementary information file. Asterisks (*) indicate statistical significance compared with control cells without JA treatment (P < 0.01, Student’s t-test). Hash (#) indicates statistical significance compared to SF3B1- or SF3B3-depleted cells following treatment with JA (P < 0.01, Student’s t-test).