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hepa_22697_sm_SupFig1.tif541KSupplemental Figure 1 Legend. Expression of exogenous human TLR4 in LX-2 or mouse TLR4 -/- HSCs. LX-2 cells or mouse TLR4-/- HSC lines were transiently or stably reconstituted with human TLR4 cDNAs. The protein expression and the localization of exogenous human TLR4 proteins following reconstitution were determined by immunopercipitation and direct immunofluorescence, respectively, using anti-Flag monoclonal antibodyies. A&B: Western blot analysis of TLR4-Flag proteins in LX-2 cells (A) and mouse stellate cell lines (B) after reconstitution of TLR4. C&D; E&F: Immunofluorescence detection of TRL4-Flag protein in LX-2 cells (C&D) and mouse HSCs (E&F) after transfection of exogenous TLR4 or control cDNAs. Cell surface localization of Flag-TLR4 protein is suggested by the appearance of TLR4 staining in LX-2 cells (C) and TLR4-/- mouse stellate cells (E) transfected with human TLR4 wild type cDNA but not in LX-2 cells (D) or mouse TLR4-/- HSCs (F) transfected with a LacZ-expressing control vector.

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