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Fig. 3. Knockdown of PLD1 leads to decreased myogenic differentiation. C2C12 cells were infected with lentiviruses expressing two different Pld1 shRNAs or the empty vector, selected by puromycin for 3 days, and then subjected to analysis by (A) quantitative RT-PCR to measure levels of Pld1 and Pld2 mRNA, and (B) western blotting for PLD1. (C) The shRNA-expressing cells were grown to confluence and induced to differentiate for 3 days, followed by immunostaining for MHC. Three independent experiments were performed in each case with similar outcomes, and representative data are shown, except for A, where the average data of all experiments are shown with the error bars representing the s.d. Student's t-test was performed to compare cells infected with Pld1 shRNA viruses and empty vector virus. *P
0.01.
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