Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Binding of C/EBPbeta to the C-reactive protein (CRP) promoter in Hep3B cells is associated with transcription of CRP mRNA.

doi: 10.4049/jimmunol.181.4.2420

Figure Lengend Snippet: FIGURE 2. C/EBP, NF-B p50, STAT3, c-Rel, and TBP bind the endogenous CRP promoter. Agarose gel of a ChIP assay performed on Hep3B cells treated with cytokines IL-1 and IL-6 for 0–15 h, as described in Materials and Methods. Abs to C/EBP, NF-B p50, STAT3, c-Rel, and TBP were used in the assays with primers flanking the CRP proximal promoter (118 to 115). The mock is a no Ab control. Input is a 1/10 dilution of total chromatin after sonication and preclearing. C/EBP, NF-B p50, and input are shown (top row). STAT3, c-Rel, and mock are shown in the middle, and TBP is shown in the bottom row. Results are representative of four experiments.

Article Snippet: Human recombinant cytokines IL-6 (206-IL) and IL-1 (201-LB) were purchased from R&D Systems.

Techniques: Agarose Gel Electrophoresis, Control, Sonication

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Binding of C/EBPbeta to the C-reactive protein (CRP) promoter in Hep3B cells is associated with transcription of CRP mRNA.

doi: 10.4049/jimmunol.181.4.2420

Figure Lengend Snippet: FIGURE 3. C/EBP binds the endogenous CRP promoter in response to cytokines. Real-time PCR of ChIP assays performed on Hep3B cells treated with cytokines IL-1 and IL-6. Zero time values were determined in each case. Subsequently three time courses were followed 30 min–6 h (n 3 assays) (a), 8–16 h (n 4) (b), and 12–36 h (n 4) (c). Abs were used in the assays with primers flanking the CRP proximal promoter (118 to 115), as described in Fig. 2. Data show C/EBP occupancy expressed as fold change after subtraction of mock and normalization to input signal (see Materials and Methods). Results are an average of three to four ex- periments, each done in duplicate. Error bar represents SD. Statistical sig- nificance of each time point compared with basal levels was determined by a one-way ANOVA and is defined. , p 0.5.

Article Snippet: Human recombinant cytokines IL-6 (206-IL) and IL-1 (201-LB) were purchased from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Binding of C/EBPbeta to the C-reactive protein (CRP) promoter in Hep3B cells is associated with transcription of CRP mRNA.

doi: 10.4049/jimmunol.181.4.2420

Figure Lengend Snippet: FIGURE 4. p50 occupancy of the CRP promoter changes modestly in the presence of cytokines. Real-time PCR of ChIP assays performed on Hep3B cells treated with cytokines IL-1 and IL-6. Zero time values were determined in each case. Subsequently three time courses were followed 30 min–6 h (n 3 assays) (a), 8–16 h (n 4) (b), and 12–36 h (n 4) (c). Abs were used in the assays with primers flanking the CRP proximal pro- moter (118 to 115), as described in Fig. 2. Data show NF-B p50 occupancy expressed as fold change after subtraction of mock and nor- malization to input signal (see Materials and Methods). Results are an average of three to four experiments, each done in duplicate. Error bar represents SD. Statistical significance of each time point compared with basal levels was determined by a one-way ANOVA at p 0.5, but the experiment had insufficient statistical power to reliably calculate p values.

Article Snippet: Human recombinant cytokines IL-6 (206-IL) and IL-1 (201-LB) were purchased from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Binding of C/EBPbeta to the C-reactive protein (CRP) promoter in Hep3B cells is associated with transcription of CRP mRNA.

doi: 10.4049/jimmunol.181.4.2420

Figure Lengend Snippet: FIGURE 5. STAT3 occupancy of the CRP promoter rises modestly in response to cytokines. Real-time PCR of ChIP assays performed on Hep3B cells treated with cytokines IL-1 and IL-6. Zero time values were deter- mined in each case. Subsequently three time courses were followed 30 min–6 h (n 3 assays) (a), 8–16 h (n 4) (b), and 12–36 h (n 4) (c). Abs were used in the assays with primers flanking the CRP proximal pro- moter (118 to 115), as described in Fig. 2. Data show STAT3 occu- pancy expressed as fold change after subtraction of mock and normaliza- tion to input signal (see Materials and Methods). Results are an average of three to four experiments, each done in duplicate. Error bar represents the SD. Statistical significance of each time point compared with basal levels was determined by a one-way ANOVA is defined. , p 0.5.

Article Snippet: Human recombinant cytokines IL-6 (206-IL) and IL-1 (201-LB) were purchased from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Binding of C/EBPbeta to the C-reactive protein (CRP) promoter in Hep3B cells is associated with transcription of CRP mRNA.

doi: 10.4049/jimmunol.181.4.2420

Figure Lengend Snippet: FIGURE 6. c-Rel and TBP occupy the CRP promoter in parallel. Real-time PCR of ChIP assays performed on Hep3B cells treated with cytokines IL-1 and IL-6. Zero time values were determined in each case. Subsequently three time courses were followed 30 min–6 h (n 3 assays) (a), 8–16 h (n 4) (b), and 12–36 h (n 4) (c). Abs were used in the assays with primers flanking the CRP proximal promoter (118 to 115), as described in Fig. 2. Data show c-Rel (solid line, Œ) and TBP (dashed line, f) occupancy expressed as fold change after subtraction of mock and normalization to input signal (see Materials and Methods). a–c, Average of three to four experiments, each done in duplicate. Error bar represents SD. d and e, Profiles from individual ChIP experiments of c-Rel and TBP promoter occupancy. Statistical significance of each time point compared with basal levels was determined by a one-way ANOVA. , p 0.5 is defined for c-Rel and , p 0.5 is indicated for TBP.

Article Snippet: Human recombinant cytokines IL-6 (206-IL) and IL-1 (201-LB) were purchased from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Binding of C/EBPbeta to the C-reactive protein (CRP) promoter in Hep3B cells is associated with transcription of CRP mRNA.

doi: 10.4049/jimmunol.181.4.2420

Figure Lengend Snippet: FIGURE 7. CRP mRNA accumulates in response to cytokines. a, Rep- resentative agarose gel of RT-PCR performed on Hep3B cells treated with cytokines IL-1 and IL-6 for the indicated times (hours). CRP mRNA levels are shown at top and -actin mRNA levels are shown at bottom. b, Average quantification of band intensity measured using ImageQuant of CRP mRNA normalized to -actin mRNA (n 4 measurements). Error bar represents SD.

Article Snippet: Human recombinant cytokines IL-6 (206-IL) and IL-1 (201-LB) were purchased from R&D Systems.

Techniques: Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Binding of C/EBPbeta to the C-reactive protein (CRP) promoter in Hep3B cells is associated with transcription of CRP mRNA.

doi: 10.4049/jimmunol.181.4.2420

Figure Lengend Snippet: FIGURE 8. Composite graph of transcription factor occupancy on the CRP promoter and CRP mRNA ac- cumulation in response to cytokines. Common time points for transcription factor occupancy from the 12– 36-h data and CRP mRNA accumulation from the 3–24-h data (12, 18, and 24 h) were plotted together. The left y-axis is the fold change above mock for the transcription factor promoter occupancy. The right y- axis is average normalized band intensity for CRP mRNA accumulation. CRP mRNA (dotted dashed gray line F), C/EBP (black line ), STAT3 (light gray line ‚), p50 (light gray line E), c-Rel (short dashed line Œ), and TBP (black dashed line f) are shown.

Article Snippet: Human recombinant cytokines IL-6 (206-IL) and IL-1 (201-LB) were purchased from R&D Systems.

Techniques:

Journal: Pharmaceuticals

Article Title: Taxifolin Prevents Cisplatin Nephrotoxicity by Modulating Nrf2/HO-1 Pathway and Mitigating Oxidative Stress and Inflammation in Mice

doi: 10.3390/ph15111310

Figure Lengend Snippet: A schematic diagram illustrating the protective effect of taxifolin against cisplatin-induced AKI. Cisplatin induces oxidative stress and activates NF-κB, leading to the release of pro-inflammatory cytokines and apoptosis. Taxifolin mitigated cisplatin-induced acute kidney injury (AKI) by upregulating Nrf2 and preventing oxidative stress, inflammation, and apoptosis.

Article Snippet: Serum BUN and creatinine were assayed using Spinreact (Girona, Spain) kits and the pro-inflammatory cytokines (TNF-α and IL-6) were determined using ELISA kits supplied by R&D Systems (Minneapolis, MN, USA).

Techniques:

Journal: Psychopharmacology

Article Title: Assessment of pioglitazone and proinflammatory cytokines during buprenorphine taper in patients with opioid use disorder

doi: 10.1007/s00213-018-4986-5

Figure Lengend Snippet: Levels of proinflammatory cytokines in CSF and plasma by treatment arm

Article Snippet: Plasma and CSF samples were analyzed in duplicate with MesoScale V-PLEX Plus (Meso Scale Discovery, MSD) Custom Proinflammatory Panel 1(IL-1β, IL-6, Il-10, TNF-α) and Custom Chemokine Panel 1 (MCP-1).

Techniques: Clinical Proteomics

Journal: The Journal of Biological Chemistry

Article Title: Human TMEM2 is not a catalytic hyaluronidase, but a regulator of hyaluronan metabolism via HYBID (KIAA1199/CEMIP) and HAS2 expression

doi: 10.1016/j.jbc.2023.104826

Figure Lengend Snippet: The effects of proinflammatory cytokines in NHDFs are abrogated by the knockdown of hTMEM2 . A , the mRNA expression of human TMEM2 and HYBID in proinflammatory cytokine-stimulated NHDFs. NHDF cells were pretreated with NTC (C) or hTMEM2 (T) siRNAs. After 24 h, the media were changed with or without the proinflammatory cytokine cocktail (1.0 ng/ml PBS, same ratio mixture of TNFα, IL-1β, and IL-6). TMEM2 and HYBID mRNA expression levels were determined by qRT-PCR after 24 h of stimulation. GUSB was used as a loading control. B , the expression levels of hHYBID and hTMEM2 proteins in NHDFs. After 48 h of stimulation with the cytokine cocktail (0.1, 0.3, and 1.0 ng/ml), the protein expression levels of hHYBID and hTMEM2 were determined via immunoblotting using specific antibodies. GAPDH was used as the loading control. C , the mRNA expression of human HAS2 in cytokine-stimulated NHDFs. The total RNAs are the same as those used in A . The HAS2 mRNA expression levels were determined by qRT-PCR. GUSB was used as a loading control. D , induction of HA in cytokine-stimulated NHDF culture medium. The concentration of HA in each medium was detected using an HA Quantification Kit. The culture medium was collected at the same time as ( B ). The control concentration (PBS) was set at 1.0 and the HA amount was shown as ratio (fold increase). A , C , and D , values represent the mean ± S.E.M. (n = 3). ∗∗ p < 0.01, ∗∗∗ p < 0.005 versus the control (PBS or C) (Tukey’s test).

Article Snippet: The Detroit 551 cells were seeded at a density of 2.5 × 10 4 cells/well in a 24-well plate and incubated with or without a mixture of the cytokines (0.1–1.0 ng/ml, TNF-α, IL-1β, and IL-6), IL-1β (1.0 ng/ml) or TGF-β (10 ng/ml) (FUJIFILM Wako Pure Chemical).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Control, Western Blot, Concentration Assay