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. 2014 Apr 2;9(4):e93257. doi: 10.1371/journal.pone.0093257

Figure 1. Functional validation of ATP6V0C knockdown.

Figure 1

Vesicular acidification following siRNA-mediated knockdown of ATP6V0C was assessed using Lysotracker Red (LTR). LTR-positive punctae were imaged in differentiated SH-SY5Y cells at 96 h following nucleofection of either Non-target siRNA control (A–C) or ATP6V0C siRNA (D–F). DMSO vehicle or 100 nM bafilomycin A1 (BafA1) was added to cells at 3 h prior to the addition of LTR. Scale bar  = 20 μm. Inset boxes in panels A and D are magnified in panels B and E respectively, with arrows indicating LTR-positive punctae. (G) Flow cytometric quantification of LTR relative mean fluorescence intensity (MFI) in vehicle-treated cells at 96 h following nucleofection with Non-target vs. ATP6V0C siRNA. Data are expressed as mean ± SEM and are represented by six independent experiments. #p<0.05 vs. Non-target siRNA using one sample t-test.