Journal: Shock (Augusta, Ga.)

Article Title: The Protective Effect of a Short Peptide Derived from Cold-inducible RNA-binding Protein in Renal Ischemia-Reperfusion Injury

doi: 10.1097/SHK.0000000000000988

Figure Lengend Snippet: Serum was collected 24 h after RIR and used to measure (A) TNF-α, (B) IL-6, (C) IL-1β using ELISA. Data are expressed as mean ± SEM (sham: n = 4 mice; RIR + vehicle: n = 5 mice; RIR + vehicle: n = 5 mice) and compared by one-way analysis of variance and Student-Newman-Keuls method. *P < 0.05 versus sham; #P < 0.05 versus vehicle. RIR, renal ischemia-reperfusion; SEM, standard error of the mean; TNF-α, tumor necrosis factor-α; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Serum TNF-α, IL-6, and IL-1β were determined with an ELISA kit, specific to mouse TNF-α, IL-6, and IL-1β (BD Biosciences, San Diego, CA).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Inflammation Research

Article Title: Viperin deficiency promotes dendritic cell activation and function via NF-kappaB activation during Mycobacterium tuberculosis infection

doi: 10.1007/s00011-022-01638-3

Figure Lengend Snippet: Viperin deficiency elicits robust cytokine, chemokine and NO production of DCs. Rsad2 −/− BMDCs were infected with H37Rv (MOI = 2) for 24 h. A Proinflammatory cytokines including IL-12p40/IL-12p70, TNF-α, IL-1β and IL-6 mRNA levels were validated by qRT-PCR and B secretion levels were detected by ELISA. C Chemokines including CXCL1, CXCL2 and CXCL10 mRNA levels were validated by qRT-PCR and D secretion levels were detected by ELISA. E iNOS mRNA level was detected by qRT-PCR and F protein expression was detected by Western blot. G NO production was detected by Griess Reagent System. A , C , E Data are presented as fold change relative to uninfected controls of Rsad2 + / + BMDCs and at least three independent experiments, with each 2–3 replicates. B , D Data are at least n = 3 independent experiments with each 2–3 replicates are shown. F Densitometric analysis was performed after Western blot analysis. β-Actin served as an internal reference. The numbers below immunoblot indicates the density ratios of (Viperin or iNOS)/β-Actin. Data are presented as fold change relative to uninfected controls of Rsad2 + / + BMDCs and are representative of three independent experiments with similar results. The ratios of protein expressions of viperin and iNOS are shown in graph. G Data are presented with the uninfected control of Rsad2 + / + BMDCs is used as 100%. A – G Data shown are the mean ± SD, T -test, * p ≤ 0.05, ** p ≤ 0.01

Article Snippet: The secretion of cytokines (IL-12p70, TNF-α, IL-1β and IL-6) and chemokines (CXCL1, CXCL2, CXCL10, CCL3 and CCL4) in the supernatant was analyzed by respective ELISA kit (MultiSciences, Hangzhou, China).

Techniques: Infection, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: Inflammation Research

Article Title: Viperin deficiency promotes dendritic cell activation and function via NF-kappaB activation during Mycobacterium tuberculosis infection

doi: 10.1007/s00011-022-01638-3

Figure Lengend Snippet: Viperin regulates DC function though activation of NF-κB p65 signaling. A Phosphorylation level of NF-κB p65 was detected by Western blot in Rsad2 −/− BMDCs infected with H37Rv (MOI = 5) for 15, 30 and 60 min. B , C , E – H Rsad2 + / + BMDCs and Rsad2 −/− BMDCs were pretreated with JSH-23 (20 uM) for 1 h, following H37Rv infection (MOI = 2) for 24 h. B iNOS mRNA level was detected by qRT-PCR. C NO production was detected by Griess Reagent System. D Rsad2 + / + and Rsad2 −/− BMDCs were pretreated with L-NAME HCI (1 mM) for 1 h, following Texas Red tagged H37Rv infection for 2 h at MOI = 5. Cells were collected and incubated with fluorescently conjugated mAb CD11c + at 4 °C for 30 min. Percentage of Texas Red positive BMDCs (CD11c + ) and MFI were analyzed by flow cytometry. E Proinflammatory cytokines including IL-12p70, TNF-α, IL-1β, and IL-6 and F chemokines including CXCL1, CXCL2, and CXCL10 secretion levels were detected by ELISA . G , H The expressions of MHC II, CD80, CD86 and CD40 on BMDCs (CD11c + ) were detected by flow cytometry and MFI were assessed. A Densitometric analysis was performed after Western blot analysis. β-Actin served as an internal reference. The numbers below immunoblot indicate the density ratios of (Viperin or p-p65/p65)/β-Actin. Data are presented as fold change relative to uninfected controls of Rsad2 + / + BMDCs and at least n = 3 independent experiments with each 2 replicates are shown. The ratios of protein expressions of Viperin and p-p65/p65 were shown in GAPDH. B Data are presented as fold change relative to uninfected control of Rsad2 + / + BMDCs and at least n = 3 independent experiments with each 2 replicates are shown. C Data are presented with the uninfected control of Rsad2 + / + BMDCs is used as 100% and at least three independent experiments, with each 2–3 replicates. D Data are presented with the 0 mM control of Rsad2 + / + BMDCs is used as 100% and are representative of three independent experiments with each 4 replicates. E , F Data are at least n = 3 independent experiments with each 2 replicates are shown. G , H Data are representative of three independent experiments with each 4 replicates. A – H Data shown are the mean ± SD, T -test, * p ≤ 0.05, ** p ≤ 0.01, ns (not significant)

Article Snippet: The secretion of cytokines (IL-12p70, TNF-α, IL-1β and IL-6) and chemokines (CXCL1, CXCL2, CXCL10, CCL3 and CCL4) in the supernatant was analyzed by respective ELISA kit (MultiSciences, Hangzhou, China).

Techniques: Activation Assay, Western Blot, Infection, Quantitative RT-PCR, Incubation, Flow Cytometry, Enzyme-linked Immunosorbent Assay