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. 2009 Feb 19;28(7):889–901. doi: 10.1038/emboj.2009.29

Figure 2.

Figure 2

(A) 293 cells were transfected with bag1, bag3 or nonsense (nons) siRNAs, as indicated. After 48 h, cells were transfected with d2GFP expression plasmid together with half the amounts of the indicated siRNAs. After additional 24 h, levels of indicated proteins were detected by immunoblot analysis. (B) 293 cells were transfected with indicated siRNAs for 48 h followed by real-time PCR analysis of BAG1 and BAG3 mRNA levels. Depicted is the mean relative expression ratio (log2) ±s.e.m. *P<0.05 and ***P<0.001 versus nons, n=3. (C) BAG1 knockdown was performed in 293 cells for 48 h. Thereafter, cells were transfected with GFP or GFP-based UPS reporter genes as indicated. After additional 24 h, levels of GFP-positive proteins were detected by western-blot analysis using a GFP antibody. (D, E) Immunoblot analysis of indicated proteins from d2HEK cells treated with different concentrations of MG132 for 5 h (D) or transfected with indicated amounts of BAG1S plasmid for 24 h (E).