Journal: iScience

Article Title: IL11 stimulates ERK/P90RSK to inhibit LKB1/AMPK and activate mTOR initiating a mesenchymal program in stromal, epithelial, and cancer cells

doi: 10.1016/j.isci.2022.104806

Figure Lengend Snippet: IL11-mediated inhibition of LKB1 occurs in the livers of mice with non-alcoholic steatohepatitis WB showing hepatic levels of p- and total ERK, P90RSK, LKB1, AMPK, mTOR, P70S6K, S6RP, ɑSMA, and GAPDH from (A) NC or WDF fed-wild-type (WT) or hepatocyte-specific Il11ra1 knockout mice (CKO). (B) NC or WDF-fed Il11ra1 +/+ (WT) and WDF-fed Il11ra1 −/− (KO) that were infected with AAV8-ALB-Null, and WDF-fed KO mice that were previously infected with AAV8-ALB-mbIl11ra1 (full-length membrane-bound Il11ra1 ) in order to specifically restore the expression of Il11ra1 in hepatocytes. (C) WDF-fed mice that were on therapeutic reversal dosing experiment; mice were treated with 10 mg/kg (2×/week; IP) of either IgG or ×209 16 weeks after the start of WDF for a duration of 8 weeks while they were still on continuous WDF feeding (n = 3 mice/group). (D) Schematic of the proposed role of ERK/P90RSK activity downstream of IL11 stimulation for the phosphorylation of LKB1 at both S325 and S428 sites, leading to LKB1 inactivation and consequent effects in stromal, epithelial, and cancer cells.

Article Snippet: Primary human tubular epithelial cells , ScienCell , Cat# 4100.

Techniques: Inhibition, Knock-Out, Infection, Membrane, Expressing, Activity Assay, Phospho-proteomics

Journal: iScience

Article Title: IL11 stimulates ERK/P90RSK to inhibit LKB1/AMPK and activate mTOR initiating a mesenchymal program in stromal, epithelial, and cancer cells

doi: 10.1016/j.isci.2022.104806

Figure Lengend Snippet:

Article Snippet: Primary human tubular epithelial cells , ScienCell , Cat# 4100.

Techniques: Virus, Plasmid Preparation, Recombinant, Protease Inhibitor, Lysis, Extraction, Activity Assay, Enzyme-linked Immunosorbent Assay, Software, Irradiation, Western Blot

Journal: Virus Research

Article Title: Transcriptomics of Marburg virus-infected primary proximal tubular cells reveals negative correlation of immune response and energy metabolism

doi: 10.1016/j.virusres.2024.199337

Figure Lengend Snippet: MARV infection of primary human proximal tubular cells (PTC). (A) MARV copies (by kallisto) at 20 h and 40 h post infection (p.i.). Mock: three uninfected PTC replicates from 40 h p.i. (B) Bright field image of mock and MARV infected PTC at 40 h p.i. taken with 10x objective at a NIKON TS-100 microscope. ( C) Principial component analysis of mock-infected, 20 h- and 40 h MARV-infected PTC, demonstrating a transcriptome difference at 20 h- and especially at 40 h p.i. (D) MARV transcriptomics at 40 h p.i.: MA plot, depicting significantly ( q <0.05) up-regulated transcripts in red and down-regulated ones in blue (cut-off fold-change ≥2). (E) Heatmap of gene expression changes after MARV infection, again demonstrating transcriptome differences at 20 h- and especially at 40 h p.i. (replicates: 1–3: mock; 4–6: 20 h p.i. and 7–9: 40 h p.i. The heatmap is color coded: blue represents down- and red upregulated genes).

Article Snippet: • Marburg virus infects and replicates in primary human proximal tubular cells (PTC).

Techniques: Infection, Microscopy, Gene Expression

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Negative regulatory responses to metabolically triggered inflammation impair renal epithelial immunity in diabetes mellitus

doi: 10.1007/s00109-012-0969-x

Figure Lengend Snippet: Renal tubular epithelial cells, but not mesangial cells, modulate overt inflammation via negative feedback mechanisms that impair immunity. a Human renal primary tubular epithelial cells ( RPTEC ) and mesangial cells ( NHMC ; Lonza) were cultured with diabetes-relevant concentrations of cytokines (TNF-α, 50 pg/ml; IL-6, 20 pg/ml; and IL-1β, 20 pg/ml), high glucose ( HG ; 20 mM) or methylglyoxal ( MG ; 10 μM) for 3 h or 5 days (120 h). Protein lysates from each condition ( n = 4) were prepared for western blot analysis using the indicated antibodies ( n ≥ 3 for each antibody). b RPTEC were first exposed to a combination of cytokines, high glucose, and methylglyoxal at concentrations stated in ( a ) for 5 days. The cells were further cultured in fresh medium in the presence of either diphenyleneiodonium chloride ( DPI ; 10 μM) or combined with Gö6983 (10 nM) for 1 day before challenge with lipopolysaccharide ( LPS ; 1 μg/ml) for 2 h. Protein lysates ( n = 4 for each condition) were analyzed by western blotting (≥3 immunoblots for each antibody). c RPTEC cultured in the combined presence of cytokines, HG and MG at the aforementioned concentrations for 5 days were challenged with a higher dose of LPS (10 μg/ml) for the indicated time points ( n = 4 for each time point). Minus 1 h denotes cells that were not exposed to LPS but were lysed for protein isolation one hour before similarly cultured cells were challenged with LPS. Protein lysates from each condition were prepared for western blotting as described (Figs. and legends). d RPTEC and NHMC were treated as follows before harvesting for protein and total RNA isolations ( n = 4 for each condition). I Untreated control cells; II incubated with cytokines (TNF-α, 50 pg/ml; IL-6, 20 pg/ml; and IL-1β, 20 pg/ml), HG (20 mM), and MG (10 μM) for 5 days; III incubated with MG (10 μM) only for 5 days; IV challenged with LPS (10 μg/ml), lipoteichoic acid ( LTA ; 10 μg/ml), and poly(I:C) (10 μg/ml) for 2 h; V treated as in ( II ) before returning to normal culture medium for 1 day and subsequently challenged with microbial stimulants as in ( IV ); VI cultured as in ( II ) before treatment with DPI (10 μM) and Gö6983 (10 nM) for 1 day in fresh culture medium; VII cultured as in ( VI ) before challenge as in ( IV ). Protein lysates were assayed using quantitative protein arrays. e Results of qPCR array plotted as fold change (RPTEC of each condition cf. untreated RPTEC). Genes with similar functions or involvement in the same biological process were grouped. A Diabetes related, B inflammatory pathways, C antioxidant system, D negative immune regulators, E interleukin and cytokine receptors, F TLR signaling, and G antimicrobial molecules and adaptive immunity. Roman numerals ( I–V ) indicate the type of treatment. I RPTEC incubated with cytokines (TNF-α, 50 pg/ml; IL-6, 20 pg/ml; and IL-1β, 20 pg/ml), HG (20 mM), and MG (10 μM) for 5 days; II RPTEC challenged with LPS (10 μg/ml), LTA (10 μg/ml) and poly I/C (10 μg/ml) for 2 h; III RPTEC treated as in ( I ), returned to normal culture medium for 1 day and then challenged with microbial analogs as in ( II ); IV RPTEC cultured as in ( I ) before combined treatment with DPI (10 μM) and Gö6983 (10 nM) for 1 day in fresh culture medium; V RPTEC cultured as in ( IV ) before being challenged as in ( II ). The qPCR array experiment for each of the conditions was performed twice

Article Snippet: Clonetics® human primary renal tubular epithelial cells (RPTEC) and mesangial cells (NHMC), purchased from Lonza BioSciences, were cultured in REGMTM Renal Epithelial Cell Growth Medium and NHMC in MsGMTM Mesangial Cell Growth Medium, respectively.

Techniques: Cell Culture, Western Blot, Isolation, Control, Incubation