Journal: Cell & Bioscience

Article Title: Biologically active, high levels of interleukin-22 inhibit hepatic gluconeogenesis but do not affect obesity and its metabolic consequences

doi: 10.1186/s13578-015-0015-0

Figure Lengend Snippet: Chronic administration of rmIL-22 protein does not affect body weight and insulin resistance in HFD-fed mice. Mice were fed a HFD for 5 months and then injected with rmIL-22 (20 ng/g body weight, i.p. injection, twice a week) or saline for an additional four weeks. a Body weights were measured. b Fasting blood glucose levels. Values represent the mean ± SEM (n = 8)

Article Snippet: To further clarify this discrepancy, we treated HFD-fed mice with rmIL-22 (R&D system) (20 ng/g body weight, twice a week) for 4 weeks.

Techniques: Injection, Saline

Journal: Cell & Bioscience

Article Title: Biologically active, high levels of interleukin-22 inhibit hepatic gluconeogenesis but do not affect obesity and its metabolic consequences

doi: 10.1186/s13578-015-0015-0

Figure Lengend Snippet: Injection of a single dose of recombinant mouse IL-22 (rmIL-22) protein reduces blood glucose levels in HFD- and streptozotocin (STZ)-treated mice. a Mice were fed a HFD for 8 weeks and then injected with saline or rmIL-22 for 2 h. Blood glucose levels were detected 120 min post IL-22 injection. b , c C57BL/6 mice were injected with STZ for 5 consecutive days. Twenty eight days later, mice were injected with saline or rmIL-22, and sacrificed 2 h later. Pancreas weights and insulin levels were measured (panel b ). Glucose levels were measured at various time points post rmIL-22 injection (panel c ). Values represent the mean ± SEM (n = 10). * P < 0.05 and ** P < 0.01 compared with the corresponding saline treated groups. d C57BL/6 mice were treated with rmIL-22 for 2 h, pancreas tissues were collected for immunostaining with anti-pSTAT3 antibody. Representative positive pSTAT3 nuclei in acinar cells are indicated by yellow arrows but not in islets (indicated by dotted lines)

Article Snippet: To further clarify this discrepancy, we treated HFD-fed mice with rmIL-22 (R&D system) (20 ng/g body weight, twice a week) for 4 weeks.

Techniques: Injection, Recombinant, Saline, Immunostaining

Journal: Cell & Bioscience

Article Title: Biologically active, high levels of interleukin-22 inhibit hepatic gluconeogenesis but do not affect obesity and its metabolic consequences

doi: 10.1186/s13578-015-0015-0

Figure Lengend Snippet: IL-22 inhibits hepatic gluconeogenesis without affecting glucose uptake in vivo . a - c Mice were fed a HFD for 8 weeks and then injected with ad-vector or ad-IL-22 for 5 days. A glucose tracer assay in vivo was performed. Glucose turnover rates and plasma glucose levels are shown (panel a ). A pyruvate tolerance test (PTT) was performed (panel b ). Real-time PCR analyses of gluconeogenic genes (panel c ). d - e C57BL/6 mice were fed a HFD for 8 weeks and then fasted for 4 h, followed by treatment with saline or rmIL-22 (1 μg/g) for 2 h. Real-time PCR analyses of gluconeogenic genes (panel d ). Two-deoxyglucose uptake experiments in vivo were performed (panel e ). Values represent the mean ± SEM (n=6-10). * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with the corresponding ad-IL-22-treated or rmIL-22-treated groups

Article Snippet: To further clarify this discrepancy, we treated HFD-fed mice with rmIL-22 (R&D system) (20 ng/g body weight, twice a week) for 4 weeks.

Techniques: In Vivo, Injection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Saline

Journal: Cell & Bioscience

Article Title: Biologically active, high levels of interleukin-22 inhibit hepatic gluconeogenesis but do not affect obesity and its metabolic consequences

doi: 10.1186/s13578-015-0015-0

Figure Lengend Snippet: Treatment with rmIL-22 protein inhibits gluconeogenesis in primary mouse hepatocytes via STAT3- and AMPK-dependent mechanisms. a Western blot analyses of IL-22-treated primary mouse hepatocytes. b Western blot analyses of IL-22- or insulin-treated hepatocytes. c Primary wild-type mouse hepatocytes with pre-treated PI3K or AMPK inhibitors, followed by IL-22 treatment. Primary STAT3KO mouse hepatocytes were also treated with IL-22. d The same experiments as those in panel C except all cells were pre-treated with Bt2-cAMP. In panels c and d , glucose production and gene expression were analyzed and normalized to 100 % in hepatocytes without IL-22 treatment in each group. Values represent the mean ± SEM (n = 4). * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with the corresponding hepatocytes without rmIL-22 treatment. # P < 0.05 and ## P < 0.01 compared with the corresponding hepatocytes from vehicle + WT mice with rmIL-22 treatment

Article Snippet: To further clarify this discrepancy, we treated HFD-fed mice with rmIL-22 (R&D system) (20 ng/g body weight, twice a week) for 4 weeks.

Techniques: Western Blot, Expressing

Journal: Molecular Oncology

Article Title: Progranulin sustains STAT 3 hyper‐activation and oncogenic function in colorectal cancer cells

doi: 10.1002/1878-0261.12552

Figure Lengend Snippet: NF ‐ kB /p65‐mediated regulation of progranulin expression in human normal and cancerous colonic cell lines. (A) Representative western blotting showing progranulin expression in HCEC ‐1 CT cells stimulated or not with IL ‐6, IL ‐22, TNF ‐α, and IL ‐17A (all used at 25 ng·mL −1 ) for 8 h. β‐actin was used as a loading control. One of four representative experiments in which similar results were obtained is shown. Right inset. Quantitative analysis of progranulin/β‐actin protein ratio in total extracts of HCEC ‐1 CT cells stimulated or not with IL ‐6, IL ‐22, TNF ‐α, and IL ‐17A for 8 h, as measured by densitometry scanning of western blots. Values are expressed in arbitrary units and are the mean ± SEM of four experiments. Differences among groups were compared using one‐way analysis of variance ( ANOVA ) followed by Tukey's post hoc test. TNF ‐α‐stimulated cells vs untreated cells, *** P < 0.001. (B) Representative western blotting showing p‐ STAT 3 Tyr705, p‐ NF ‐ kB /p65 Ser536, p‐p38, and p‐ ERK expression in HCEC ‐1 CT cells stimulated or not with IL ‐6, IL ‐22, TNF ‐α, and IL ‐17A (all used at 25 ng·mL −1 ) for 15 min (left panels), 30 min (middle panels), and 8 h (right panels). β‐actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. (C) Representative western blotting showing progranulin, p‐ NF ‐ kB /p65 Ser536, p‐p38, and p‐ ERK expression in HCEC ‐1 CT cells either left untreated or stimulated with TNF ‐α (used at 25 ng·mL −1 ) for 8 h in the presence of NF ‐ kB /p65 inhibitor ( BAY 11‐7082), p38 inhibitor ( SB 202190), ERK inhibitor ( PD 98059), or DMSO (vehicle) as indicated. β‐actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. (D). Representative western blotting showing progranulin, p‐ NF ‐ kB /p65 Ser536, p‐p38, and p‐ ERK expression in HT ‐29 cells cultured for 8 h in the presence of NF ‐ kB /p65 inhibitor ( BAY 11‐7082), p38 inhibitor ( SB 202190), ERK inhibitor ( PD 98059), or DMSO (vehicle) as indicated. β‐actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown.

Article Snippet: Freshly isolated TILs were resuspended and cultured in complete RPMI 1640 medium and cell‐free supernatants harvested after 48 h. To investigate whether IL‐6, IL‐22, TNF‐α, and IL‐17A modulate progranulin expression as well as STAT3, NF‐kB/p65, p38, and ERK activation, HCEC‐1CT cells were stimulated with recombinant human IL‐6 (Peprotech, London, UK), IL‐22 (R&D Systems, Minneapolis, MN, USA), TNF‐α (R&D Systems), or IL‐17A (Peprotech) (all used at 25 ng·mL −1 ) for 0.25–24 h. Progranulin expression and STAT3, NF‐kB/p65, p38, and ERK activation were assessed by western blotting.

Techniques: Expressing, Western Blot, Cell Culture