Extended Data Fig. 2: Normal maturation and integrin cell surface expression of MCs upon TLN-1 depletion.
From: Slow integrin-dependent migration organizes networks of tissue-resident mast cells
(a) Scheme for mouse strains and procedures to analyze TLN-1 function in several types of MCs, including bone marrow-derived MCs (BMMCs, cultured with additional IL-4) and peritoneal MCs (PMCs). (b, c) The efficiency of conditional TLN-1 knockout was confirmed by immunoblot analysis (b) and flow cytometry (c) by using a pan-talin antibody (recognizing TLN-1 and TLN-2). Cell lysates were generated from BMMC or PMC cultures of WT and Mcpt5-Cre+/− Tln1fl/fl mice. Actin was used as loading control. Representative images of two biological replicates (b). Intracellular flow cytometry of BMMCs, gated by FSC/SSC, confirmed the results from the immunoblot. Secondary antibody control is shown on top (grey filled). (d) Normal expression of the MC maturation markers c-KIT and FcεR1 in BMMC cultures of WT and Mcpt5-Cre+/− Tln1fl/fl mice. Gating for BMMC by FSC/SSC in (d) was done as in (c). (e) Normal cell surface expression of integrin subunits on Tln1−/− BMMCs (in comparison to WT BMMCs, dataset from Extended Data Fig. 1a). (f) Normal β1 and β2 integrin cell surface expression in endogenous skin MCs of Mcpt5-Cre+/− Tln1fl/fl Rosa26LSL:TdTomato (Tln1ΔMC) mice in comparison to MCs from Mcpt5-Cre+/− Tln1+/fl Rosa26LSL:TdTomato (WT) mice. Quantification of flow cytometric measurements for integrin expression was based on n (WT) = 3 and n (Tln1ΔMC) = 3 mice. Bars display the mean; P = 0.9361 (β1), P = 0.2217 (β2), ns, non-significant, two-sided t tests.
