Fig. 5. PLD1 does not function through the actin cytoskeleton in C2C12 differentiation. C2C12 cells were infected with lentiviruses expressing two different Pld1 shRNAs or a scrambled hairpin sequence as a negative control, selected with puromycin for 3 days and induced to differentiate. Separately, the non-infected cells were subjected to 0.5% 1- or 2-butanol treatment during differentiation. At 0, 12 hours and 24 hours differentiation, the cells were fixed and labeled with Rhodamine-phalloidin. The samples to be compared were stained simultaneously, with image capture performed with an identical exposure time.