Fig 2: Expression of transcription factors involved in initiation of valve development in Tie2Cre;Piezo1cKO mice. (A and B) FOXC2 (A) or NFATc1 (B) with Prox1 staining in E17.5 mesentery of WT or Tie2Cre;Piezo1cKO embryos. Insets show magnified images. FOXC2: n = 8 for WT, n = 7 for Tie2Cre;Piezo1cKO embryos. NFATc1: n = 7 for WT, n = 6 for Tie2Cre;Piezo1cKO embryos. (Scale bar: 100 µm.)

Fig 3: NK1R-signaling Promotes Activation of NFAT1, NFAT2, and NF?b-p65, and T Cell Survival Dependent on IL-2 Secretion(A) Calcineurin activity in WT and NK1RKO T cells incubated with SarSP or CD3 Ab, measured 15 min after treatment. Means ± 1 SD of duplicated results of 1 representative experiment of 2.(B) Western blot analysis of NFAT1, NFAT2, NF?ß-p65, cFos, and cJun, in nuclear extracts of WT and NK1RKO T cells, untreated or after incubation with CD3 and CD28 Ab. Controls were stimulated with ionomycin (Iono). One representative experiment of 3.(C) Concentrations of IL-2 by ELISA in supernatants of WT and NK1RKO T cells untreated or after 24-h stimulation with CD3 and CD28 Ab. Means ± 1 SD, 1 representative of 3 experiments.(D) Surface IL-2Ra expression by FACS in WT and NK1RKO T cells untreated or after 24-h stimulation with CD3 and CD28 Ab. One representative of 3 experiments.(E) Comparison by FACS of T cell proliferation (CFSE dilution), cell death (FVD incorporation), and activation (CD44High) between WT and NK1RKO T cells after 4-day stimulation with CD3 and CD28 Ab. Numbers are cell percentages per quadrant. One representative out of 6 experiments.(F) Comparison by FACS of cycles of cell division (histograms) and percentages of cell proliferation (bar diagram) between WT and NK1RKO T cells. Numbers in histograms are cell percentages. Each dot in the bar diagrams represents a mouse. Means ± 1 SD.(G) Percentages of cell death (by FVD incorporation) of WT and NK1RKO T cells stimulated for 4 days with CD3 and CD28 Ab, alone or plus IL-2. Means ± 1 SD, 6 mice per condition.Results were analyzed by 1-way ANOVA followed by ad hoc Student Newman Keuls test (A, C, F, and G). ***p < 0.001, ****p < 0.0001, NS: not significant.