Journal: The Journal of Clinical Investigation
Article Title: FOXO1 pathway activation by VISTA immune checkpoint restrains pulmonary ILC2 functions
doi: 10.1172/JCI184932
Figure Lengend Snippet: ( A ) Overview of VISTA downstream signaling. ( B – E ) Freshly isolated WT and Vsir –/– aILC2s were stained intranuclearly for p-AKT, FOXO1, p65, and p52. Representative histograms of protein expression of p-AKT ( B ), FOXO1 ( C ), p65 ( D ), and p52 ( E ), and corresponding quantifications as MFI. ( F – I ) WT aILC2s were treated with FOXO1 inhibitor or vehicle. ( G and H ) Representative plots of GATA-3 ( G ) and Ki67 ( H ) expression levels and corresponding quantification (as MFI). ( I ) Cytokine levels in cell supernatant (per 10 3 ILC2s). ( J – M ) WT aILC2s were treated with FOXO1 activator or vehicle. ( K and L ) Representative plots of GATA-3 ( K ) and Ki67 ( L ) expression levels and corresponding quantification (as MFI). ( M ) Cytokine levels in cell supernatant (per 10 3 ILC2s). Data are presented as mean + SEM and are representative of at least 2 independent experiments. Statistical significance was assessed using 2-tailed Student’s t test; ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Article Snippet: When specified, 10 μg/mL VISTA agonist antibody (MH5A, BioLegend), 10 μg/mL VISTA antagonist antibody (13F3, Bio X Cell), 100 nM FOXO1 inhibitor (AS1842856, MedChemExpress), or 1 μM FOXO1 activator (LOM612, MedChemExpress) with corresponding isotype control or vehicle (DMSO) was further added to cultures for the indicated times.
Techniques: Isolation, Staining, Expressing