Anti rabbit cy3 cy5

Livers were fixed in 4% paraformaldehyde (Polysciences, Inc., Warrington, PA) and embedded in paraffin or Tissue-Tek® Optimum Cutting Temperature (OCT) Sakura Finetek, Torrance, CA) for sectioning. Immunofluorescence (IF) staining signals for PROM1 (1:50 dilution, eBiosciences, San Diego, CA), CYTOKERATIN-19 (1:100 dilution, gift from JR Friedman, University of Pennsylvania, Philadelphia. PA), and LacZ (1:400, Bioss, Woburn, MA) were detected by secondary antibodies conjugated either with anti-mouse cyanine (Cy) 3, Cy5, anti-rat Cy3/Cy5, or anti-rabbit Cy3/Cy5 (1:200 dilution, Jackson ImmunoResearch Labs, West Grove, PA) (9 (link), 10 (link), 20 (link)). Images were then acquired by a Leica DM5500B IF microscope using Leica Suite Advanced Fluorescence (LAS AF) 6000 software (Leica Microsystems, Wetzlar, Germany). Immunohistochemistry was performed using 2.1 μg/mL anti-INTEGRIN-β6 (gift from Shelia Violette, Biogen, Inc). Bright-field images were captured using a Leica DM1000 (DFC290) transmitted light microscope (Leica Microsystems AG, Heerbrugg, Switzerland) with Leica Application Suite (version 2.7.1R1).

Livers were fixed in 4% paraformaldehyde (PFA, Poly Sciences Inc., Warrington, PA), embedded in paraffin and then sectioned. Immunofluorescence imaging was performed as previously described [12 (link)] using primary antibodies listed in Supplemental Table 1 and secondary antibodies conjugated either with anti-mouse Cy3/Cy5, anti-rat Cy3/Cy5, or anti-rabbit Cy3/Cy5 (Jackson Immuno Research Lab, West Grove, PA). Images were acquired using a Leica DM5500B immunofluorescence microscope with Leica Suite Advanced Fluorescence (LAS AF) 6000 software (Wetzlar, Germany) or a LSM700 confocal system with ZEN software (Carl Zeiss Microimaging, Thornwood, NY). Bright field images were acquired using a Leica DM1000 (DFC290) microscope interfaced with Leica Application Suite, Version 2.7.1R1 (Leica Microsystems, Switzerland). Sirius red staining was performed as previously described [9 ].

Livers were fixed in 4% paraformaldehyde (PFA, Poly Sciences Inc., Warrington, PA) and embedded in paraffin for sectioning. Immunofluorescence staining was performed as described previously (9 (link)) (Supplemental Table 1). Signals were detected by secondary antibodies conjugated either with anti-mouse Cy3/Cy5, anti-rat Cy3/Cy5, or anti-rabbit Cy3/Cy5 (Jackson Immuno Research Lab, West Grove, PA). Fluorescence images were acquired by an LSM700 confocal system controlled by ZEN software (Carl Zeiss Microimaging, Thornwood, NY) or by a Leica DM5500B immunofluorescence microscope using Leica Suite Advanced Fluorescence (LAS AF) 6000 software (Wetzlar, Germany). Bright field images were acquired using a Leica DM1000 (DFC290) transmitted light microscope (Leica Microsystems, Switzerland) using Leica Application Suite, Version 2.7.1R1.