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. 2013 Jun 19;8(6):e67216. doi: 10.1371/journal.pone.0067216

Figure 10. Hsp60 siRNA increases N-SMase activity in N-SMase2–overexpressed HEK293 cells.

Figure 10

HEK293 cells were seeded into six-well dishes, and 24 h later, they were transfected with the negative control (mock and scramble) or Hsp60 siRNA (20 nM). After 48 h, the cells were transfected with N-SMase2 for 24 h. Total protein was extracted, and the N-SMase activity was determined (A) followed by immunoblotting for Hsp60, N-SMase2, and GAPDH (B). The results represent the mean ± S.D. of five independent experiments. *p<0.05 compared with scramble control. After siRNA transfection for 48 h, the cells were transfected with N-SMase2 and incubated for the indicated time periods. Total protein was extracted, and N-SMase activity was determined (C) followed by immunoblotting for Hsp60, N-SMase2, and GAPDH (D). These results represent the mean ± S.D. from a single experiment performed in triplicate. Similar results were obtained in three independent experiments.