Phospho-NFAT2 (Ser172) Antibody - #AF8293

Fig. 4 11R-VIVIT inhibits the nuclear localization and dephosphorylation of NFAT2 in RTECs in an AKI-to-CKD progression model. a Double immunofluorescence staining of NFAT2 (green) and DAPI (blue) and merged images of kidneys from the sham-operated, IRI and IRI + 11R-VIVIT treatment groups. Scale bars = 20 μm. b Quantification of the percentage of renal tubular epithelial cells with NFAT2 expression in the nucleus. *P 

Figure 6 Role of Orai1-mediated store-operated Ca2+ entry (SOCE) in parathyroid hormone (PTH)-induced NFAT nuclear translocation in human umbilical vein endothelial cells (HUVECs). (A–F) Representative confocal microscopy images and summary data (I) showing NFATC1 distribution in HUVECs treated for 24 h with (A) vehicle control, (B) 100 pM PTH, (C) PTH + CsA (calcineurin inhibitor), (D) PTH + W7 (calmodulin antagonist), (E) PTH + BTP2 (an Orai nonspecific inhibitor) or (F) PTH + Orai1 siRNA transfection. Green fluorescence indicates NFATC1; Blue, 4′,6-diamidino-2-phenylindole (DAPI) indicates nuclei. Data are shown as the mean ± SEM; n = 4. **P < 0.01, ***P < 0.001 vs. Con or PTH analyzed by one-way analysis of variance followed by Dunnett's multiple comparisons test. (G,H) Representative Western blot images showing fractionation assay results indicating the presence of p-NFATC1 in the cytoplasmic [Cyto, (H)] and NFATC1 in the nuclear [Nuc, (G)] extracts under the same treatment conditions as for confocal microscopy analyses. Lamin B1 is a nuclear marker; β-Tubulin is a cytoplasmic marker. (I) Summary data showing the ratio of green fluorescence intensity of NFATc-GFP in the nuclear (Nuc)/cytoplasmic (Cyto).

Figure 6 Role of Orai1-mediated store-operated Ca2+ entry (SOCE) in parathyroid hormone (PTH)-induced NFAT nuclear translocation in human umbilical vein endothelial cells (HUVECs). (A–F) Representative confocal microscopy images and summary data (I) showing NFATC1 distribution in HUVECs treated for 24 h with (A) vehicle control, (B) 100 pM PTH, (C) PTH + CsA (calcineurin inhibitor), (D) PTH + W7 (calmodulin antagonist), (E) PTH + BTP2 (an Orai nonspecific inhibitor) or (F) PTH + Orai1 siRNA transfection. Green fluorescence indicates NFATC1; Blue, 4′,6-diamidino-2-phenylindole (DAPI) indicates nuclei. Data are shown as the mean ± SEM; n = 4. **P < 0.01, ***P < 0.001 vs. Con or PTH analyzed by one-way analysis of variance followed by Dunnett's multiple comparisons test. (G,H) Representative Western blot images showing fractionation assay results indicating the presence of p-NFATC1 in the cytoplasmic [Cyto, (H)] and NFATC1 in the nuclear [Nuc, (G)] extracts under the same treatment conditions as for confocal microscopy analyses. Lamin B1 is a nuclear marker; β-Tubulin is a cytoplasmic marker. (I) Summary data showing the ratio of green fluorescence intensity of NFATc-GFP in the nuclear (Nuc)/cytoplasmic (Cyto).

FIGURE 6 | Role of Orai1-mediated store-operated Ca2+ entry (SOCE) in parathyroid hormone (PTH)-induced NFAT nuclear translocation in human umbilical vein endothelial cells (HUVECs).(G,H) Representative Western blot images showing fractionation assay results indicating the presence of p-NFATC1 in the cytoplasmic

Figure 6. NIFE Inhibits the Expression of PD-1 on T Cells in an NFAT2-Dependent Manner In Vitro and In Vivo (A) Left panel: the expression of PD-1 in peripheral blood mononuclear cells (PBMCs) treated with NIFE was detected by qPCR. Right panel: the expression of PD1 in PBMCs transfected with NFAT2 siRNA was detected by qPCR. (B) Upper panel: the transcriptional regulation of PD-1 by NFAT2 and p-STAT3 was detected by ChIP. Lower panel: the expression of p-NFAT2 and CAMK-IV in the indicated PBMCs was detected by WB. (C) The binding sites between NFAT2 and the PD-1 promoter were confirmed by a dual-luciferase reporter system. (D) The relationship between NFAT2 and PD-1 in human T cells was detected from the GEO database (GEO: GSE56580). (E) The percentages of PD-1+ T cells (upper panels) and PD-1+ CD8+ T cells (lower panels) in peripheral T cells from CRC patients treated with NIFE were detected by flow cytometry sorting. (F) The percentages of PD-1+ T cells (upper panels) and PD-1+ CD8+ T cells (lower panels) in peripheral T cells from healthy people treated with NIFE and cocultured with SW620 cells were detected by flow cytometry sorting.

Figure 7. NMU regulates ILC2 activation via PI3K/NFAT and MEK/NFAT signals. ILC2s were sorted from the lungs of mice on day 3 after RSV infection, and co-cultured with NMU in vitro in the presence or absence of signal inhibitors. A-C Expressions of IL-5 and IL-13 by Real-time PCR in the presence of MEK inhibitor U0126-EtOH (A), PI3K inhibitor LY294002 (B), and NFAT inhibitor 11R-VIVIT (C). D-E Concentrations of IL-5 and IL-13 in the culture supernatants of pulmonary ILC2s were determined by ELISA in the presence of MEK inhibitor U0126-EtOH (D), PI3K inhibitor LY294002 (E), and NFAT inhibitor 11R-VIVIT (F). G-H The expression of signal proteins in the isolated ILC2s were determined by Western blot under the conditions of the presence or absence of U0126-EtOH (G) or LY294002 (H). Data are representative of at least two individual experiments, error bars represent SEM;