Journal: The Journal of Cell Biology

Article Title: FGF-23–Klotho signaling stimulates proliferation and prevents vitamin D–induced apoptosis

doi: 10.1083/jcb.200803024

Figure Lengend Snippet: Signaling events induced by FGF-23–Klotho. Immunoblotting showing that FGF-23 or Klotho alone have no effect on kinase activity in PTEC or FHs74Int cells. Combined effects of FGF-23 and Klotho show increased phosphorylation of ERK1/2, p38, JNK, AKT, IκB, and GSK-3β. α-Tubulin was used as a loading control.

Article Snippet: Human renal PTEC were obtained from Clontech Laboratories, Inc. FHs74Int human small intestine epithelial cells were acquired from the American Type Culture Collection.

Techniques: Western Blot, Activity Assay, Phospho-proteomics, Control

Journal: The Journal of Cell Biology

Article Title: FGF-23–Klotho signaling stimulates proliferation and prevents vitamin D–induced apoptosis

doi: 10.1083/jcb.200803024

Figure Lengend Snippet: FGF-23–Klotho prevents vitamin D–induced apoptosis. (A) ELISA analysis of 1α-hydroxylase expression showing no significant changes in PTEC cells exposed to FGF-23 or Klotho alone but greatly decreased levels when exposed to both FGF-23 and Klotho. Small molecule inhibitors against Ras and PI3K were sufficient to provide marginal rescue of this decrease in expression. No significant changes were found for treatment of FHs74Int cells. Graphs represent mean ± SD ( n = 3). *, P < 0.05. (B) Flow cytometry analysis for TUNEL staining of cells exposed to exogenous vitamin D showing that it caused extremely high levels of apoptosis. Addition of FGF-23 and Klotho was sufficient to rescue most of the vitamin D–induced apoptosis, whereas FGF-23 or Klotho alone did not. PI3K inhibitor prevented this rescue, whereas Ras inhibitor had no effect. (C) ELISA for active caspase-3 levels, showing the same patterns as observed with the TUNEL analysis. Graphs represent mean ± SD ( n = 3). *, P < 0.001.

Article Snippet: Human renal PTEC were obtained from Clontech Laboratories, Inc. FHs74Int human small intestine epithelial cells were acquired from the American Type Culture Collection.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, TUNEL Assay, Staining

Journal: Clinical immunology (Orlando, Fla.)

Article Title: CVID Enteropathy is Characterized by Exceeding Low Mucosal IgA Levels and Interferon-Driven Inflammation Possibly Related to the Presence of a Pathobiont

doi: 10.1016/j.clim.2018.09.008

Figure Lengend Snippet: Interferons Drive Inhibition of Lipid Metabolism in Epithelial Cells. (a) Expression of IFNB1 and IFN type I dependent genes (RSAD2, OAS2, MX1) in duodenal biopsies from of CVID patients with (red), without (blue) enteropathy and control subjects (green), all genes passed false discovery rate <0.03 comparing E-CVID to other two groups. (b) expression of IFNG and IFN type II dependent genes (GBP1, IDO, TAP1); (c) Gene expression ratios between E-CVID and control individuals (y axis) and between interferon treated versus untreated human epithelial cells (x axis; n=3 per group), left panel both types of interferons; middle panel interferon type I; right panel interferon type II. (d) Expression of genes defined as lipid metabolism by Gene Ontology and downregulated in E-CVID, representative genes are indicated; left panel: median values in E-CVID and noE-CVID tissues; right panel: individual values in cells treated and untreated with both interferons, red is high and blue is low expression; (e) intracellular cholesterol levels in epithelial cells treated or untreated with both types of interferons data presented as mean±s.e.m.; each column number of biological replicates equals 14 (**p<0.0001 by one-tailed Wilcoxon test); (f) model of CVID enteropathy.

Article Snippet: Human epithelial cell line (FHs 74 Int) was acquired from ATCC and maintained according to ATCC protocol using Hybri-Care Medium (cat# ATCC® 46-XTM) with addition of 10% FBS (ATCC, cat# 30–2020, lot: 62144240), 5% Penicillin Streptomycin (Corning, REF 30–002-CI) and 30 ng/ml epidermal growth factor (EGF; Tonbo Biosciences, cat# 21–8356-M001).

Techniques: Inhibition, Expressing, Control, Gene Expression, One-tailed Test