Journal: International Journal of Molecular Sciences

Article Title: Moderate Physical Activity as a Prevention Method for Knee Osteoarthritis and the Role of Synoviocytes as Biological Key

doi: 10.3390/ijms20030511

Figure Lengend Snippet: ( A – E ): IL-1β immunohistochemistry. ( A ) IL-1β immunolabeling was very weak in the synovium of Group 1; ( B ) IL-1β immunolabeling was very weak in the synovium of Group 2; ( C ) IL-1β immunolabeling was moderate in the synovium of Group 3; ( D ) IL-1β immunolabeling was weak in the synovial membrane of Group 4; ( E ) Graph representing the densitometric count (Log2 densitometric count − pixel 2 ) of IL-1-immunolabeling identified among groups. For details, see the text. ( F – J ): IL-4 immunohistochemistry. ( F ) IL-4 immunolabeling was moderate in the synovium of Group 1; ( G ) IL-4 immunolabeling was strong in the synovium of Group 2; ( H ) IL-4 immunolabeling was weak in the synovium of Group 3; ( I ) IL-4 immunolabeling was strong in the synovial membrane of Group 4; ( J ) Graph representing the densitometric count (Log2 densitometric count − pixel 2 ) of IL-4-immunolabeling identified among groups. For details, see the text. ( K – O ): IL-6 immunohistochemistry. ( K ) IL-6 immunolabeling was moderate in the synovium of Group 1; ( L ) IL-6 immunolabeling was strong in the synovium of Group 2; ( M ) IL-6 immunolabeling was very strong in the synovium of Group 3; ( N ) IL-6 immunolabeling was very strong in the synovial membrane of Group 4; ( O ) Graph representing the densitometric count (Log2 densitometric count − pixel 2 ) of IL-6-immunolabeling identified among groups. For details, see the text. ( P – T ): IL-10 immunohistochemistry. ( P ) IL-10 immunolabeling was moderate in the synovium of Group 1; ( Q ) IL-10 immunolabeling was strong in the synovium of Group 2; ( R ) IL-10 immunolabeling was weak in the synovium of Group 3; ( S ) IL-10 immunolabeling was strong in the synovial membrane of Group 4; and, ( T ) Graph representing the densitometric count (Log2 densitometric count − pixel 2 ) of IL-10-immunolabeling identified among groups. For details, see the text. In inserts are the image analyses by the software in which red color represents immunolabeling. ( A – D , F – I , K – N , P – S ): Objective lens, 20×; scale bars: 50 µm. Results were presented as the mean ± SD. ANOVA was used to evaluate the significance of the results. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; ns, not significant.

Article Snippet: After blocking, the sections were incubated overnight at 4°C with rabbit polyclonal anti-IL-1β (ab9787; Abcam), diluted 1:100 in PBS (Bio-Optica); rabbit polyclonal anti-IL-4 (ab9622; Abcam), diluted 1:100 in PBS (Bio-Optica); goat polyclonal anti-IL-6 antibody (sc-1265; Santa Cruz Biotechnology Inc., Dallas, TX, USA) diluted 1:100 in PBS (Bio-Optica); rabbit polyclonal anti-IL-10 (ab34843; Abcam), diluted 1:100 in PBS (Bio-Optica); mouse monoclonal anti-TNF-α antibody (ab199013; Abcam ) diluted 1:100 in PBS (Bio-Optica); mouse monoclonal anti-MMP-13 antibody (sc-81547; Santa Cruz Biotechnology Inc.) diluted 1:100 in PBS (Bio-Optica); rabbit polyclonal anti-lubricin antibody (ab28484; Abcam), diluted 1:100 in PBS (Bio-Optica).

Techniques: Immunohistochemistry, Immunolabeling, Software

Journal: International Journal of Molecular Sciences

Article Title: Novel Medicinal Mushroom Blend as a Promising Supplement in Integrative Oncology: A Multi-Tiered Study using 4T1 Triple-Negative Mouse Breast Cancer Model

doi: 10.3390/ijms21103479

Figure Lengend Snippet: Primary/secondary antibodies and respective dilution used for immunocytochemical experimental procedures.

Article Snippet: Primary antibodies , Anti-Interleukin-6 (M-19) , Purified antibody raised against a peptide mapping at the C-terminus of IL-6 of mouse origin , Santa Cruz Biotechnology (Santa Cruz, CA, USA), Goat polyclonal IgG, Cat# sc-1265, RRID: AB_2127470 , 1:100.

Techniques: Purification, Plasmid Preparation

Journal: The Journal of Biological Chemistry

Article Title: Inducible microRNA-590-5p inhibits host antiviral response by targeting the soluble interleukin-6 (IL6) receptor

doi: 10.1074/jbc.RA118.005057

Figure Lengend Snippet: miR-590-5p regulates antiviral signaling through sIL6R. A, luciferase assays of IL6R−/− A549 cells cotransfected with IFN-β, IFN-λ1, NF-κB, and ISRE luciferase reporter constructs and pRL-TK together with mimic control (miR-Ctrl) or miR-590-5p mimics and infected with SeV (m.o.i. = 1) for 12 h. Rel. Lucif. Act., relative luciferase activity. B, qRT-PCR analysis of IFN-α, IFN-β, and IFN-λ1 mRNA levels in IL6R−/− A549 cells transfected with mimic control or miR-590-5p mimics and then infected with SeV for 8 h. C, immunoblot analysis of mIL6R and sIL6R in IL6R+/+ and IL6R−/− A549 cells reconstituted with empty vector, sIL6R, mIL6R, or sIL6R and mIL6R together. The intensities of mIL6R and sIL6R were normalized to β-actin. D–F, qRT-PCR analysis of IFN-β (D), IFN-λ1 (E), and IL6 (F) mRNA levels in IL6R−/− A549 cells reconstituted with empty vector, sIL6R, mIL6R, or sIL6R and mIL6R together; transfected with mimic control or miR-590-5p mimics; and then infected with SeV for 8 h. G and H, IL6R−/− A549 cells reconstituted with empty vector, sIL6R, mIL6R, or sIL6R and mIL6R together were transfected with miR-590-5p mimics (G), miR-590-5p inhibitor (H), or their controls and then infected with IAV (m.o.i. = 1) for 12 h. Relative levels of NP-specific mRNA, cRNA, and vRNA were measured by qRT-PCR. All experiments were repeated at least three times with similar results. Error bars represent S.D. **, p < 0.01; ***, p < 0.001; n.s., not significant (analysis of two-way ANOVA followed by Bonferroni post-test).

Article Snippet: Antibodies (Abs) against β-actin (60008-1-Ig) and mIL6R (23457-1-AP) were purchased from the ProteinTech Group (Wuhan, China).

Techniques: Luciferase, Construct, Infection, Activity Assay, Quantitative RT-PCR, Transfection, Western Blot, Plasmid Preparation

Journal: The Journal of Biological Chemistry

Article Title: Inducible microRNA-590-5p inhibits host antiviral response by targeting the soluble interleukin-6 (IL6) receptor

doi: 10.1074/jbc.RA118.005057

Figure Lengend Snippet: miR-590-5p down-regulates endogenous sIL6R and mIL6R expression. A and B, qRT-PCR analysis of sIL6R (A) and mIL6R (B) mRNA levels in A549 cells transfected with mimic control (miR-Ctrl) or miR-590-5p mimics and infected with IAV (m.o.i. = 1) for the indicated time. C, ELISA analysis of sIL6R protein in the cell culture supernatants harvested in A549 cells treated as described in A and B. D, immunoblot analysis of mIL6R and sIL6R protein in A549 cells treated as described in A and B. The intensities (Rel. Intensit.) of mIL6R and sIL6R were normalized to β-actin (graphs below). E and F, qRT-PCR analysis of sIL6R (E) and mIL6R (F) mRNA levels in A549 cells transfected with miR-590-5p inhibitor or miRNA inhibitor control and infected with IAV (m.o.i. = 1) for the indicated time. G, ELISA analysis of sIL6R protein in the cell culture supernatants harvested in A549 cells treated as described in E and F. H, immunoblot analysis of mIL6R and sIL6R protein in A549 cells treated as described in E and F. The intensities of mIL6R and sIL6R were normalized to β-actin (graphs below). All experiments were repeated at least three times with similar results. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s., not significant (analysis of two-way ANOVA followed by Bonferroni post-test).

Article Snippet: Antibodies (Abs) against β-actin (60008-1-Ig) and mIL6R (23457-1-AP) were purchased from the ProteinTech Group (Wuhan, China).

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