Journal: Japanese Clinical Medicine

Article Title: Dynamic Change in Cells Expressing IL-1β in Rat Hippocampus after Status Epilepticus

doi: 10.4137/jcm.s13738

Figure Lengend Snippet: Figure 2. IL-1b immunoreactivity in the hippocampus of control rat without SE and after SE. (A, D, G, J, M) Expression of IL-1b at control rat hippocampus. (A) Expression of IL-1b at the hippocampus of control rat without SE in low-power magnification. Scale bar = 1 mm. (D) CA1 region of control rat in intermediate power magnification. (G) CA3 region of control rat in intermediate power magnification. D, G; scale bar = 100 μm. (J) CA1 region of control rat in high-power magnification. (M) CA3 region of control rat in high-power magnification. J, M; scale bar = 50 μm. Expression of IL-1b was faintly observed in the pyramidal cells at CA1 (D, J) and CA3 (G, M) regions. (B, E, H, K, N) Expression of IL-1b at day 1 after SE. (B) Expression of IL-1b at the hippocampus at day 1 after SE in low power magnification. Scale bar = 1 mm. (E) CA1 region at day 1 after SE in intermediate power magnification. (H) CA3 region at day 1 after SE in intermediate power magnification. E, H; scale bar = 100 μm. (K) CA1 region at day 1 in high power magnification. (N) CA3 region at day 1 after SE in high power magnification. IL-1b expression increased transiently in the cytoplasm of the remaining pyramidal cells in the CA3 region beginning on day 1 after SE (H, N). Expression of IL-1b in the cytoplasm of pyramidal cells at CA3 was greater than that at CA1 on day 1 after SE (E, H, K, N; Table 1). K, N; scale bar = 50 μm. (C, F, I, L, O) Expression of IL-1b at day 21 after SE. (B) Expression of IL-1b at the hippocampus at day 21 after SE in low-power magnification. Scale bar = 1 mm. (E) CA1 region at day 21 after SE in intermediate power magnification. (H) CA3 region at day 21 after SE in intermediate power magnification. F, I; scale bar = 100 μm. (F) CA1 region at day 21 in high power magnification. (N) CA3 region at day 21 after SE in high power magnification. Reactive astrocyte-like cells emerging in CA1 showed IL-1b immunoreactivity on day 7. This immunointensity increased in proportion to progressive hypertrophy until day 21. L, O; scale bar = 50 μm.

Article Snippet: For the immunohistochemical study, a commercially available rabbit polyclonal anti-IL-1b antibody (diluted 1:500; sc-1251, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used.

Techniques: Control, Expressing

Journal: Japanese Clinical Medicine

Article Title: Dynamic Change in Cells Expressing IL-1β in Rat Hippocampus after Status Epilepticus

doi: 10.4137/jcm.s13738

Figure Lengend Snippet: Figure 3. Double-label immunofluorescence staining of IL-1b and GFAP. Co-localization of IL-1b (red, A) and GFAP (green, B) is shown with an immunofluorescence method. Co-localization is visualized in yellow in the merged image (C). Double-label fluorescent immunohistochemistry clarified that reactive astrocytes expressed IL-1b. Scale bar = 50 μm.

Article Snippet: For the immunohistochemical study, a commercially available rabbit polyclonal anti-IL-1b antibody (diluted 1:500; sc-1251, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used.

Techniques: Immunofluorescence, Staining, Immunohistochemistry

Journal: Japanese Clinical Medicine

Article Title: Dynamic Change in Cells Expressing IL-1β in Rat Hippocampus after Status Epilepticus

doi: 10.4137/jcm.s13738

Figure Lengend Snippet: Figure 4. Total IL-1b expression in the hippocampus after SE. The total expression level IL-1b in the hippocampus measured by using Luminex technology was significantly elevated from day 1 after SE and maintained till day 21 (P 0.01). Asterisks (*) and daggers (†) indicate significant differences (P 0.05 and P 0.005, respectively) from the value for the control group.

Article Snippet: For the immunohistochemical study, a commercially available rabbit polyclonal anti-IL-1b antibody (diluted 1:500; sc-1251, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used.

Techniques: Expressing, Luminex, Control

Journal: Frontiers in immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits.

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: FIGURE 3 Inhibition of TLR2 signaling pathway and NF-kB activation by novel-miR1. (A) qPCR results demonstrate the reduction of TLR2, TNF-a, IL-1b, and IL- 6 mRNA levels by novel-miR1. Each test was performed in triplicate. Fold change was calculated based on qPCR results. (B) Western blot assay results indicate the decrease in TLR2, P65, pP65, TNF-a, IL-1b, and IL-6 protein levels by novel-miR1. (C) Western blot assay results showing the decrease in protein levels of TLR2, P65, pP65, TNF-a, IL-1b, IL-6 (fold change was calculated by the gray values of the image). ns, p ≥0.05; *, p ≤ 0.05; ***, p ≤0.001.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-kB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-kB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit antiTNF-a polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1b monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-b-actin monoclonal antibody (1:2,000 dilution, Beyotime TABLE 1 Primer sequences used in quantitative PCR.

Techniques: Inhibition, Activation Assay, Western Blot

Journal: Frontiers in immunology

Article Title: Cysticercus pisiformis -derived novel-miR1 targets TLR2 to inhibit the immune response in rabbits.

doi: 10.3389/fimmu.2023.1201455

Figure Lengend Snippet: FIGURE 5 C. pisiformis inhibits the TLR2 signaling pathway and NF-kB activation. (A) Expression profile of novel-miR1 in rabbit PBLCs over a 3-month period. (B) Relative expression TLR2 mRNA in rabbit PBLCs during the initial 3-month period. (C) Western blot assay results indicate inhibition of TLR2, P65, and pP65 protein levels in C. pisiformis-infected rabbit PBLCs. (D) Western blot assay results demonstrating the inhibition of TLR2, P65, and pP65 protein levels in C. pisiformis-infected rabbit PBLCs (the fold change was calculated by the gray values of the image). (E) ELISA assay results indicate inhibition of IL-6, TNF-a, and IL-1b protein expression levels in C. pisiformis-infected rabbit PBLCs. *, p ≤0.05; **, p ≤0.01; ***, p ≤0.001.

Article Snippet: The separated proteins were transferred to a PVDF membrane and blocked with 5% skimmed milk at room temperature for 3 h. Rabbit anti-TLR2 monoclonal antibody (1:1,000; Abcam; cat. no. Ab209217), phospho-NF-kB p65 (Ser536) rabbit monoclonal antibody (1:1,000; Abcam; cat. no. ab239882), NF-kB p65 rabbit polyclonal antibody (1:1,000; Abcam; cat. no. Ab16502), rabbit anti-IL-6 polyclonal antibody (1:1,000; Bioss, Woburn, MA, USA; cat. no. bs-0782R), rabbit antiTNF-a polyclonal antibody (1:1,000; Bioss; cat. no. bs-2081R), mouse anti-IL-1b monoclonal antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA; cat. no. sc52012), and mouse anti-b-actin monoclonal antibody (1:2,000 dilution, Beyotime TABLE 1 Primer sequences used in quantitative PCR.

Techniques: Activation Assay, Expressing, Western Blot, Inhibition, Infection, Enzyme-linked Immunosorbent Assay