Journal: PLoS ONE

Article Title: Upregulation of sphingosine-1-phosphate receptor 3 on fibroblast-like synoviocytes is associated with the development of collagen-induced arthritis via increased interleukin-6 production

doi: 10.1371/journal.pone.0218090

Figure Lengend Snippet: (A) Development of collagen-induced arthritis in WT and S1P 3 knockout (S1P 3 -KO) mice. Arthritis was evaluated three times per week during Days 21–42. Disease severity was measured by assessing the clinical score of all four paws, in which the maximum possible score for an individual mouse was 16. Statistical analysis was performed at Days 36, 38, 40, and 42. Results are presented as the mean ± SEM (n = 20 mice per group) and were analyzed by t -test with Bonferroni’s correction. * P < 0.05 and ** P < 0.01 (WT vs. KO mice). (B) Synovial inflammation, bone erosion, and cartilage destruction were examined histologically on H&E-stained sections of ankle joints and assigned scores of 1–4. Results are presented as box-and-whisker plots (n = 20 mice per group) and were analyzed by Mann-Whitney U-test. The middle hash, the box, and the whiskers represent the median, interquartile range (IQR), and 10/90 percentile values respectively. * P < 0.05, NS = not significant. (C) Representative images of H&E-stained ankle joint sections are shown at a magnification of 40×. Left panel: WT specimen assigned a score of 3 for all parameters. Right panel: KO specimen assigned a score of 1 for inflammation and 0 for the other parameters. (D) Anti-CII IgG 1 and IgG 2 antibodies in serum from CIA mice were measured in an ELISA. Results are presented as box-and-whisker plots (n = 10 mice per group) and were analyzed by Mann-Whitney U-test. The middle hash, the box, and the whiskers represent the median, IQR, and minimum/maximum values respectively. (E) Expression of TNFα and IL-6 in arthritic synovium was analyzed by real-time RT-PCR. Results are presented as the mean ± SEM (n = 6 mice per group) and were analyzed by t -test. * P < 0.05, NS = not significant.

Article Snippet: Levels of anti-CII IgG 1 and IgG 2 were measured using a Mouse Anti-type II Collagen IgG Subtype Antibody ELISA Kit (Chondrex), according to the manufacturer’s instructions.

Techniques: Knock-Out, Staining, Whisker Assay, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: Diabetes

Article Title: Endogenous oils derived from human adipocytes are potent adjuvants that promote IL-1α-dependent inflammation.

doi: 10.2337/db13-1476

Figure Lengend Snippet: Figure 1—Endogenous oils have potent adjuvant properties. A: Female C57BL/6 mice were immunized intraperitoneally on day 0 with PBS or OVA (200 mg/mouse) either alone or mixed with an equal volume (100 mL) of endogenous oils derived from the human omentum. OVA was also mixed with an equal volume (100 mL) of FIA, and this solution was emulsified by sonication. The mice were boosted with these same treatments on day 14 and were killed on day 21. Blood samples were recovered and the serum was removed following centrifugation. OVA-specific antibody titres (IgG, IgG1, IgG2b, and IgG2c) in the serum were determined by ELISA. Results are mean titres (+SEM) for five mice per group. Versus PBS (OVA): *, P < 0.05; **, P < 0.01; ***, P < 0.001. B: Female C57BL/6 mice were immunized intraperitoneally with PBS or OVA (200 mg/mouse) either alone or mixed with an equal volume (100 mL) of endogenous oils or an aqueous or organic fraction derived from this preparation following a lipid-extraction procedure. OVA was also mixed with an equal volume (100 mL) of FIA, and this solution was emulsified by sonication. The mice were boosted with these same treatments on day 14 and were killed on day 21. Blood samples were recovered and the serum was removed following centrifugation. OVA-specific antibody titres (IgG, IgG1, IgG2b,, and IgG2c) in the serum were determined by ELISA. Results are mean titres (+SEM) for four mice per group. Versus PBS (OVA), **, P < 0.01; ***, P < 0.001. Organic fraction (OVA) versus aqueous fraction (OVA): ccc, P < 0.001. n.s., not significant.

Article Snippet: Measurement of Antigen-Specific Serum Antibodies by ELISA Titres of antigen-specific total serum IgG as well as the IgG subtypes IgG1, IgG2b, and IgG2c were determined using commercially available antibodies from SigmaAldrich (total IgG), BD Pharmingen (IgG1 and IgG2b), and AbD Serotec (IgG2c).

Techniques: Adjuvant, Derivative Assay, Sonication, Centrifugation, Enzyme-linked Immunosorbent Assay, Extraction

Journal: eNeuro

Article Title: Excitatory Amino Acid Transporter EAAT5 Improves Temporal Resolution in the Retina

doi: 10.1523/ENEURO.0406-21.2021

Figure Lengend Snippet: Primary antibodies and markers

Article Snippet: The 20F12 antibody (rat IgG 2 subtype) was detected with secondary mouse anti-rat-IgG2 antibody (gift of R. Feederle, Helmholtz Zentrum München; 1:100 in CTA; 1 h at RT) and visualized with donkey-anti-mouse Cy3 antibody.

Techniques: