Characterizing Immune Cell Subsets in Mouse Intestine

Lamina propria cells from SI of ΔdblGata or WT mice were stained with phycoerythrin (PE)- conjugated anti-c-Kit, fluorescein isothiocyanate (FITC)-conjugated anti-β7 integrin, Horizon V500-conjugated CD4, APC-Cy-7-conjugated anti-CD3e (BD Pharmingen, Mountain View, CA, USA), allophycocyanin (APC)-conjugated anti-IL-17RB, PE-Cy7-conjugated anti-FcεRIα (Biolegend, San Diego, CA, USA) and biotinylated anti-T1/ST2. Subsequently, cells were counterstained with PerCP-Cy5.5-conjugated monoclonal antibodies against lineage (Lin) markers (CD11b, CD11c, CD45R (BD Pharmingen, Mountain View, CA, USA) CD8α, Ly6G, and Pacific Blue labelled Streptavidin (Biolegend, San Diego, CA, USA)) before analysis with a FACS Canton II (BD Bioscience, Mountain View, CA, USA). CD4⁺ Th₂ cells were identified as Lin^-, CD3⁺, CD4⁺, and IL17RB⁺ populations. ILC2 cells were identified as Lin^-, CD3^-, CD4^-, 1L17RB⁺, and c-Kit^- populations. MMC9 cells were identified as Lin^-, CD3^-, CD4^-, 1L17RB^-, c-Kit⁺, and FcεRIα⁺ populations as previously described [38 (link)]. All cytometric data was acquired using BD FACSCanto II and data analysis was performed using Flowjo software (FlowJo, Ashland, OR, USA).