Fig. 4.

Validation of differential protein expression. A, Western blotting of eight differentially expressed proteins (FBL, STAT5B, MGMT, VIM, VASP, GNAI2, C1QBP and PSMD10) at 48 h after HOTAIR inhibition. Values are expressed as the percent change relative to the controls and Western blots are from one representative experiment. Western blot analysis detected changes in expression consistent with those detected by MS. GAPDH was used as an internal control. The change in the expression of VIM and FBL was further verified by MRM. MS spectrum of target peptides of VIM (B) and FBL (C) selected for MRM validation. Results of the MRM analysis were consistent with SILAC data.