Validation of the LSECs Tfr1 knockout in the Tfrc^Tek-Cremouse model. (A) Frozen liver slices from control Tfrc^fl/fl (left) or Tfrc^Tek-Cre mice (right) were processed for immunofluorescence and stained for Tfr1 using a Cy3-labeled secondary antibody (red) and for CD31 using an Alexa488-labeled secondary antibody (green). Overlapping areas of Tfr1 and CD31 expressions are shown in yellow. Nuclei were visualized using 4′,6-diamidino-2-phenylindole (DAPI) staining (blue). Areas in the highlighted rectangles are shown at a higher magnification (bottom). Arrows indicate exclusive CD31 expression in the LSEC lining. Scale bars, 10 μm (highlighted rectangles, bottom, scale bars, 2 μm). (B) Frozen liver slices from a Rosa26^mT-mG/+;Tfrc^fl/+;Tek-Cre reporter mouse were processed for confocal microscopy imaging for the expression of mT (red) and mG (green); the latter emerged following Cre-mediated recombination. The arrows indicate the LSEC lining. Scale bars, 10 μm.