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. 2019 Jul 15;129(8):3310–3323. doi: 10.1172/JCI127166

Figure 7. CASTOR1 and CASTOR2 inhibit pre–miR-K1 and -K4–induced cell proliferation.

Figure 7

(A–C) CASTOR1 and CASTOR2 inhibited pre–miR-K1 and -K4–induced mTORC1 activation. ΔmiR-mutant cells stably expressing vector control (ΔmiR-V) or pre–miR-K4 (ΔmiR–pre–miR-K4 in A and B) or pre–miR-K1 (ΔmiR–pre–miR-K1 in C) were transduced with increasing doses of lentiviruses of CASTOR1, CASTOR2, or vector control at 2, 4, or 6 MOI for 48 hours and examined by Western blotting for expression of the mTORC1 downstream effectors p-S6K and p-4EBP1. Three independent experiments were repeated with similar results, and results from one representative experiment were shown. (D) CASTOR1 and CASTOR2 inhibited pre–miR-K4 and -K4–induced cell proliferation. ΔmiR-mutant cells stably expressing a vector control (ΔmiR-V), pre–miR-K4 (ΔmiR–pre–miR-K4) or pre–miR-K1 (ΔmiR–pre–miR-K1) were transduced with increasing doses of lentiviruses of CASTOR1, CASTOR2, or vector control at 2, 4, or 6 MOI for 48 hours and examined for cell proliferation. Three independent experiments were repeated with similar results, and results from 1 representative experiment with 4 biological replicates are shown as the mean ± SEM. (E and F) CASTOR1 and CASTOR2 inhibited pre–miR-K4 and -K4–induced cell-cycle progression and induced weak apoptosis. ΔmiR-mutant cells stably expressing vector control (ΔmiR-V), pre–miR-K4 (ΔmiR–pre-miR-K4) or pre–miR-K1 (ΔmiR–pre–miR-K1) were transduced with increasing doses of lentiviruses of CASTOR1, CASTOR2, or vector control at 2, 4, or 6 MOI for 48 hours and examined for cell-cycle progression (E) and apoptosis (F). Three independent experiments were repeated with similar results, and results from 1 representative experiment with 3 biological replicates are shown as the mean ± SEM. Data were analyzed by 1-way ANOVA followed by Tukey’s post hoc test for P values below 0.05. *P < 0.05, **P < 0.01, and ***P < 0.001.