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. 2015 Aug 26;10(8):e0136313. doi: 10.1371/journal.pone.0136313

Fig 1. IDN5706 reduces the levels of EDEM1 in a time-dependent manner.

Fig 1

(A) H4 cells were left untreated or treated for different periods of time with 250 μM IDN5706, and subsequently analyzed by Western blotting with an EDEM1 antibody. The antibody specificity was validated on protein extracts from cells stably expressing either luciferase shRNA, used as positive control (shLuc), or EDEM1 shRNA (shEDEM1). Specific antibodies against ER proteins including Calnexin, Gp78 and Bip were tested. (B) Densitometric quantification of the levels of EDEM1 in cells left untreated or treated with IDN5706 for 16 h, as shown in A. Bars represent the mean ± SD of three independent experiments (***, P < 0.001). (C) H4 cells were either left untreated (lane 1) or transfected with a plasmid encoding HA-epitope-tagged EDEM1 (lanes 2 and 3). After 16 h cells were left untreated or treated with 250 μM IDN5706 for 8 h, and cellular extracts were subjected to SDS-PAGE followed by immunoblot with mouse antibody anti-HA-epitope. In A and C, Western blotting with antibody to β-actin was used as loading control. The position of molecular mass markers is indicated on the left.