A. Representative flow cytometric acquisition plot showing
gated LSEC preparation from BALB/c mouse showing two color flow cytometric
analysis of 2.4G2 FITC plus F4/80 PE, and histogram representation of single
Fluor expression. The results are representative of 4 different experiments and
mice/biological replicates. B. Five color flow cytometric analysis
of gated LSEC from BALB/c mouse for 2.4G2 PE-Cy7, F4/80 PE, NK-1.1 FITC, Desmin
Pac-blue, and Cd11c Percp. Two colors analyzed at one given time are shown in
one Figure. C. The picture shows the SEM image of LSEC from a
BALB/c mouse. Arrows point fenestrae. The insert projected represents zoomed
view. D. Representative two color flow cytometric analysis of gated
LSEC from 2B-KIX mouse for 2.4G2 FITC plus 4F5 594 along with histograms showing
single colors from 3 different experiments and mice. In A-D, events from isotype
controls are represented as blue dots/blue peaks and from primary antibodies as
red dots/red peaks. The average percentage of events showing single or double
positive expression from three different LSEC preparations is indicated in
respective quadrants. E. Confocal microscopic image of LSEC from
2BKIX showing labeling of 4F5 (top row) and 2.4G2 (bottom row) along with DIC
and DAPI (nucleus). F. The picture shows the SEM image of LSEC from
a 2B-KIX mouse. Arrows point fenestrae. Note folded and layered cell membranes
of LSEC. The scale bar in C, E and F indicates 5μm.