Proteins were extracted from 106 cultured LX‐2 cells. For this purpose, T‐PER Tissue Protein Extraction Reagent as lysis buffer supplemented with proteinase and phosphatase inhibitors (Thermo Fisher Scientific) was used. The protein concentration was measured by protein assay (Bio‐Rad), and all samples were normalized to 100 μg. Western blotting was performed as described previously. 27 The membranes were incubated overnight with antibodies against ERK1/2 (#9102, Cell Signaling Technology), phospho‐ERK1/2 (#4370S), Akt (#4691), phospho‐Akt (#4060), SMAD2/3 (#3102), phospho‐SMAD2/3 (#8828), PDGFRα (#3174), phospho‐PDGFα (Tyr1018) (#4547), PDGFRβ (#3169), phopho‐PDGFRβ (Tyr751) (#3166) and β‐actin (#4967). Densitometric analysis was performed with ImageJ software version 64 (National Institutes of Health).

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