Mouse primary IECs were isolated from wild-type
Dhx15fl/fl and
Dhx15IEC-KO mice. The
cells were then fixed and stained with APC/Cyanine7 anti-mouse CD45 antibody
(30-F11, Biolegend), PE anti-mouse CD324 (E-Cadherin) antibody (DECMA-1,
Biolegend), PE/Cyanine7 anti-mouse CD326 (EpCAM) antibody (G8.8, Biolegend)
and their isotype matched control antibodies for their differentiation.
Cells isolated from spleen, mesenteric lymph node (LN), and lamina propria
lymphocytes were stained using live/dead Zombie Aqua Fixable viability Kit
(Biolegend) for 10 minutes followed by staining with fluorochrome-conjugated
antibodies on ice for 20 minutes, washed twice in PBS/BSA, and fixed in 1%
paraformaldehyde prior to flow cytometry analysis. Flow cytometry data were
acquired on a LSR-II flow cytometer (Beckton Dickinson) and analyzed using
FlowJo v10 software (Tree Start) as previously described (Xing et al., 2016 (link)).
Dhx15fl/fl and
Dhx15IEC-KO mice. The
cells were then fixed and stained with APC/Cyanine7 anti-mouse CD45 antibody
(30-F11, Biolegend), PE anti-mouse CD324 (E-Cadherin) antibody (DECMA-1,
Biolegend), PE/Cyanine7 anti-mouse CD326 (EpCAM) antibody (G8.8, Biolegend)
and their isotype matched control antibodies for their differentiation.
Cells isolated from spleen, mesenteric lymph node (LN), and lamina propria
lymphocytes were stained using live/dead Zombie Aqua Fixable viability Kit
(Biolegend) for 10 minutes followed by staining with fluorochrome-conjugated
antibodies on ice for 20 minutes, washed twice in PBS/BSA, and fixed in 1%
paraformaldehyde prior to flow cytometry analysis. Flow cytometry data were
acquired on a LSR-II flow cytometer (Beckton Dickinson) and analyzed using
FlowJo v10 software (Tree Start) as previously described (Xing et al., 2016 (link)).