Fig. 3: Lrig1-deficiency in Treg cells impairs the suppressive activity.
a The level of Lrig1 expression on the cell surface of iTreg cells whose Lrig1 expression was knocked down by 50 nM or 150 nM of 2 different siRNAs targeting Lrig1 (siLrig1) or 150 nM of scrambled sequences (siControl) (n = 3). Data are expressed as mean ± S.E.M (***P < 0.001 versus siControl-1, ###P < 0.001 versus siControl-2). b Quantification of the suppressive activity of Lrig1-silenced iTreg cells (n = 3) Data are expressed as mean ± S.E.M (***P < 0.0001). c siLrig1- or siControl- nucleofected naïve CD4+ T cells from Foxp3-IRES-GFP mice were induced to differentiate into iTreg cells, and the level of Foxp3 in CD4+Foxp3+ (GFP+) cells was examined. d Representative images (left), and quantification (right) of the suppressive potential of iTreg cells from Lrig1+/+, Lrig1+/−, or Lrig1−/− mice (n = 3). Data are expressed as mean ± S.E.M (***P < 0.001, ++P = 0.005, +++P < 0.001, ##P = 0.002, ###P < 0.001, $$P = 0.002, &&&P < 0.001, @@@P < 0.001). * symbol shows versus 1:1 of Lrig1+/+, + symbol shows versus 1:2 of Lrig1+/+, # symbol shows versus 1:4 of Lrig1+/+, $ symbol shows versus 1:1 of Lrig1+/−, & symbol shows versus 1:2 of Lrig1+/−, @ symbol shows versus 1:4 of Lrig1+/−. e Representative dot plots of the level of IFNγ, IL-4, IL-17A, or Foxp3 in each cell type differentiated from Lrig1+/− or Lrig1−/− mice. f Mouse splenocytes were isolated from Lrig1+/− or Lrig1−/− mice, and the level of IFNγ+, IL-4+, IL-17A+, or Foxp3+ in CD4+ T cells was examined (n = 9). Data are expressed as mean ± S.E.M (IFNγ: P = 0.9341, IL-4: P = 0.1721, IL-17A: P = 0.0934, Foxp3: P = 0.2866). Ordinary one-way ANOVA with Dunnett’s multiple comparisons test (a, b), two-way ANOVA with Tukey’s multiple comparisons test (d) or two-tailed unpaired Student’s t test (f). All experiments were repeated at least three times. Source data are provided as a Source Data file.
