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Figure 6


Fig. 6. PLD1 regulates the mTOR-IGF2 pathway during C2C12 differentiation. (A) C2C12 cells were infected with lentiviruses expressing two different Pld1 shRNAs, the empty vector or a scrambled hairpin sequence (Scram), selected with puromycin, and then induced to differentiate for 3 days. The cell lysates were analyzed by western blotting. (B) Total RNA was extracted from cells expressing Pld1 shRNAs or treated with 0.5% 1-butanol or 50 nM rapamycin, and Igf2 mRNA levels were measured by quantitative RT-PCR. (C) IGF2 protein levels in the conditioned media from cells in A were analyzed by ELISA. (D) Cells stably expressing H19-luc-ME were infected with lentiviruses as above, induced to differentiate for 3 days, and then subjected to luciferase assays. For B-D, the average results of three independent experiments are shown, with error bars representing the s.d. Student's t-test was performed to compare PLD1 shRNA with the negative control (vector or Scram). *P<0.05; **P<0.01.





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