Schematic representation of SILAC-based proteomic approach on membrane proteins of differentiating RLSCs. (A) Phase contrast of RLSC and RLSCdH, magnification 20X. (B) Western blot analysis in total cell extract for the indicated proteins. For each fraction, 10 μg of protein sample were loaded; Hepatocyte nuclear factor 4 (HNF4) was used to assess cellular differentiation of RLSCs into RLSCdH; and GAPDH was used as proteins loading control. One representative experiment out of three is shown. (C) Whole cell extract (WCE), soluble (S) and membrane (M) fractions of mixed RLSC and DH protein samples were analyzed by western blotting. E-cadherin and GAPDH were used respectively as membrane and soluble markers to assay the membrane protein purification.