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

Journal: PLoS ONE

Article Title: Iron-Ascorbate-Mediated Lipid Peroxidation Causes Epigenetic Changes in the Antioxidant Defense in Intestinal Epithelial Cells: Impact on Inflammation

doi: 10.1371/journal.pone.0063456

Figure Lengend Snippet: Cells were incubated with Fe/Asc (200 µM/2 mM) and Trolox (0.25 mM) for 6 h at 37°C and/or with 5-AZA (10 µM). The protein expression of Interleukin-6 (IL-6) (A, C) and cyclooxygenase 2 (COX2) (B, D) was determined by western blot as described in Materials and Methods. Results represent the means ± SEM of n = 3 independent experiments. *P<0.05, **P<0.01 vs. controls; # P<0.05, ## P<0.01 vs. Fe/Asc.

Article Snippet: The different primary antibodies were added as follow: 1∶1000 rabbit anti-COX-2 (74 kDa; Novus, Oakville, ON), 1∶5000 goat anti-NF-κB (65 kDa; Santa Cruz Biotechnology, Santa Cruz, CA), 1∶5000 rabbit anti-IκBα (39 kDa; Cell signaling, Beverly, MA) and 1∶500 mouse anti-IL-6 (25 kDa; R&D Systems, Minneapolis, MN).

Techniques: Incubation, Expressing, Western Blot