Anti phospho nfat5 ser145 antibody

Samples were incubated with anti-F4/80 antibody (Cell Signaling Technology, Cat# 70076) or anti-phospho-NFAT5 (Ser145) antibody (Invitrogen, Cat# PA5-105436) and anti-IL-20 antibody (7E) at 4 °C overnight. Next day, the sections were incubated for 1 h with Alexa Fluor®488-conjugated anti-mouse secondary antibody (Jackson ImmunoResearch Labs, Cat# 115-545-003) and Alexa Fluor®594-conjugated anti-rabbit secondary antibody (Jackson ImmunoResearch Labs, Cat# 111-585-003), and finally mounted with DAPI (Vector Laboratories, Cat# H-1200). Pictures were taken under an inverted fluorescence microscope IX71 (Olympus).

Cells were incubated with anti-BAX antibody (Biolegend, Cat# 633602) or anti-BCL-2 antibody (Biolegend, Cat# 633502) or anti-phospho-NFAT5 (Ser145) antibody (Invitrogen, Cat# PA5-105436) or anti-IL-20 antibody (7E) or anti-IL-20R1 antibody (Abcam, Cat# ab90935) or anti-IL-20R2 antibody (Abcam, Cat# ab95824) or anti-IL-22R1 antibody (Abcam, Cat# ab5984) at 4 °C overnight. The next day, the sections were washed with PBS and incubated with the secondary antibody for 1 h. Reactions were detected using AEC chromogen stain (ScyTek Laboratories, Cat# ACG500) and nuclei were counterstained with hematoxylin (ScyTek Laboratories, Cat# HMM500).

Thirty micrograms of protein were loaded into SDS-PAGE gel (10 or 15%) and ran for 3 h. Protein gel was transferred to a 0·45 µm PVDF membrane and was blocked by 5% milk or 5% BSA dissolved in TBST. Antibodies used in this study include anti-NFAT5 antibody (Invitrogen, Cat# PA1-023), anti-phospho-NFAT5 (Ser145) antibody (Invitrogen, Cat# PA5-105436), anti-IL-20 antibody (7E), and anti-GAPDH antibody (Proteintech, Cat#, 10494-1-AP). The original western blot images are provided (Additional file 2).