Journal: Biomolecules

Article Title: Intestinal Barrier in Post- Campylobacter jejuni Irritable Bowel Syndrome

doi: 10.3390/biom13030449

Figure Lengend Snippet: EEA1 and cytokine intensity grade (0–4) at surface cells in the colon mucosa of PI-IBS patients and controls.

Article Snippet: Briefly, 3 µm paraffin sections were deparaffinized followed by retrieval of the antigenic epitopes via treatment in a pressure cooker in citrate buffer pH 6 for 3 min. Unspecific bindings were blocked by a serum-free protein block (DAKO, Glosdrup, Denmark) for 15 min at room temperature and the following polyclonal primary antibodies—rabbit-anti-human-EEA1 (abcam, Cambridge, UK), IL-1β (Bioss, Woburn, MA, USA), IL-22 (abcam), IL-4 (Tebubio, Frankfurt, Germany), IL-10 and IFNγ (both Peprotech, Hamburg, Germany), and goat-anti-human-IL-6 (R&D systems, Minneapolis, MN, USA) and -TNFα (PeproTech)—were applied in dilutions previously titrated in antibody diluent (DAKO) at 4 °C overnight.

Techniques:

Journal: Biomolecules

Article Title: Intestinal Barrier in Post- Campylobacter jejuni Irritable Bowel Syndrome

doi: 10.3390/biom13030449

Figure Lengend Snippet: Immunohistochemistry of Th1 cytokines (red) in the colon mucosa of controls and PI-IBS patients of IL-1β ( a , b ), IL-6 ( c , d ), IL-22 ( e , f ), INFγ ( g , h ) and TNFα ( i , j ), (counterstaining with hematoxylin = violet). Staining for IL-1β and IL-22 intensified in surface enterocytes in PI-IBS. Bar = 100 µm.

Article Snippet: Briefly, 3 µm paraffin sections were deparaffinized followed by retrieval of the antigenic epitopes via treatment in a pressure cooker in citrate buffer pH 6 for 3 min. Unspecific bindings were blocked by a serum-free protein block (DAKO, Glosdrup, Denmark) for 15 min at room temperature and the following polyclonal primary antibodies—rabbit-anti-human-EEA1 (abcam, Cambridge, UK), IL-1β (Bioss, Woburn, MA, USA), IL-22 (abcam), IL-4 (Tebubio, Frankfurt, Germany), IL-10 and IFNγ (both Peprotech, Hamburg, Germany), and goat-anti-human-IL-6 (R&D systems, Minneapolis, MN, USA) and -TNFα (PeproTech)—were applied in dilutions previously titrated in antibody diluent (DAKO) at 4 °C overnight.

Techniques: Immunohistochemistry, Staining

Journal: bioRxiv

Article Title: An AhR-Ovol1-Id1 regulatory axis in keratinocytes promotes skin homeostasis against atopic dermatitis

doi: 10.1101/2024.01.29.577821

Figure Lengend Snippet: (A) RT-qPCR results of Ovol1 and Il1a expression in epidermal cells of the HDM/SEB-treated mice at day 7. n = 3 mice per group. (B) Experimental design for IL-1R Ab treatment in (C-K). Samples were harvested at day 11 after treatment for downstream analysis. IgG was used as a control. (C-D) Flow cytometry analysis of dermal γδT cells in the back skin of IgG- or IL-1R Ab-treated SSKO mice. See legends for additional details. n = 4 mice per group. (E) Representative photographs of IgG- or IL-1R Ab-treated SSKO mice. (F) Clinical scores of IgG- or IL-1R Ab-treated SSKO mice. n = 5 mice per group. (G) Representative skin histology (H/E staining). Scale bar = 100 μm. (H) Quantification of epidermal, dermal and total skin thickness in IgG- or IL-1R Ab-treated SSKO mice. n = 4 mice per group. (I) TEWL values of IgG- or IL-1R Ab-treated SSKO mice. n = 5 mice per group. (J-K) Flow cytometry analysis of the indicated cell types in the back skin of IgG- or IL-1R Ab-treated SSKO mice. See legends for additional details. n = 4 mice per group. (L) RT-qPCR results of Il4 , Il17a and Il33 expression in whole skin. Data are mean + SEM. n = 5 mice per group. * p < 0.05, ** p < 0.01, *** p < 0.001. p values were calculated using 2-tailed unpaired Student t test or Mann-Whitney U test.

Article Snippet: For IL-1R blocking, mice were i.p injected with 200 μg IL-1R antibody (BioXcell, Cat# BE0256, Clone JAMA-147) or IgG (BioXcell, Cat# BE0091) at days −1, 0, 3, 7 and 10 of HDM/SEB treatment.

Techniques: Quantitative RT-PCR, Expressing, Flow Cytometry, Staining, MANN-WHITNEY

Journal: bioRxiv

Article Title: An AhR-Ovol1-Id1 regulatory axis in keratinocytes promotes skin homeostasis against atopic dermatitis

doi: 10.1101/2024.01.29.577821

Figure Lengend Snippet: Related to . (A-B) Percentages of different immune cell populations out of total skin cells (A) or skin CD45 + cells (B) in IgG- or IL-1R Ab-treated SSKO mice at day 11. n = 4 mice per group. (C) Clinical scores of skin eruption, scaling, bleeding and redness at day 11. n = 5 mice per group. (D) Quantification of dermal and total skin thickness. n = 4 mice per group. (E) Quantification of the indicated immune cell types per gram of skin tissue at day 11. n = 4 mice per group. (F) RT-qPCR analysis of the indicated genes in whole skin at day 11. n = 5 mice per group. (G) Quantification of the indicated immune cell types per gram of skin tissue in DMSO- or AGX51-treated SSKO mice at day 11. n = 4 mice per group. (H-I) Percentages of different immune cell populations out of total skin cells (H) or skin CD45 + cells (I) at day 11. n = 4-5 mice per group. (J) RT-qPCR results of the indicated genes in the whole skin at day 11. Data are mean + SEM. n = 4 mice per group. * p < 0.05, ** p < 0.01. p values were calculated using 2-tailed unpaired Student t test.

Article Snippet: For IL-1R blocking, mice were i.p injected with 200 μg IL-1R antibody (BioXcell, Cat# BE0256, Clone JAMA-147) or IgG (BioXcell, Cat# BE0091) at days −1, 0, 3, 7 and 10 of HDM/SEB treatment.

Techniques: Quantitative RT-PCR

Journal: International Journal of Nanomedicine

Article Title: Effectiveness of Microwave Therapy Combined with Berberine /GelMA via COX-2/IL-1β Pathway to Treat Skeletal Muscle Injury: An in vivo Study in Rats

doi: 10.2147/IJN.S500490

Figure Lengend Snippet: ( A and B ) Expression levels of mRNA for IL-1β, COX-2 in skeletal muscle of SD rats in each group. ( C and D ) The expression levels of IL-1β, PGE2 in the serum of SD rats in each group at 1/4/7 days after modeling. ( E ) Expression levels of IL-1β, COX-2 in skeletal muscle of SD rats in each group. (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).

Article Snippet: PGE2 and IL-1β Elisa kit were purchased from Abmart (China).

Techniques: Expressing

Journal: PLOS ONE

Article Title: MICT ameliorates hypertensive nephropathy by inhibiting TLR4/NF-κB pathway and down-regulating NLRC4 inflammasome

doi: 10.1371/journal.pone.0306137

Figure Lengend Snippet: SHR-M group and SHR-H group performed exercise training with different exercise prescriptions for 8 weeks. (A) The weight of rats was measured every week. (B) Systolic blood pressure, (C) Diastolic blood pressure, (D) Mean arterial pressure and (E) Heart rate, were measured after 8-week exercise training. (F) Serum creatinine (Scr), (G) Blood urea nitrogen (BUN), and (H) Urine protein were measured to evaluate the renal function. *p< 0.05, **p< 0.01, ***p< 0.001.

Article Snippet: The primary antibodies used were anti-Collagen I (1:1000; Zenbio), anti-Collagen Ш (1:1000; Huabio), TGF-β1 (1:1000; Zenbio), α-SMA (1:1000; Zenbio), anti-TLR4 (1:1000; Proteintech), anti-NF-κB P65 (1:1000; Zenbio), anti-P-P65 (1:500; Zenbio), anti-IкBα (1:1000; Zenbio), and anti-p-IкBα (1:500; Zenbio), anti-NLRC4 (1:1000; AiFang), anti-Caspase-1 (1:1000; Zenbio), anti-ASC (1:1000; Zenbio), anti-IL-1β (1:500; Huabio), anti-GAPDH (1:50000; Proteintech).

Techniques:

Journal: PLOS ONE

Article Title: MICT ameliorates hypertensive nephropathy by inhibiting TLR4/NF-κB pathway and down-regulating NLRC4 inflammasome

doi: 10.1371/journal.pone.0306137

Figure Lengend Snippet: (A) Western blot was used to detect the protein expressions of (B) NLRC4, (C) Caspase-1, (D) ASC and (E) IL-1β in WKY group, SHR-S group, SHR-M group, SHR-H group. ELISA detection of (F) IL-6 and (G) TNF-α (H) IL-1β for evaluating the concentration of pro-inflammatory factors in serum. Data are presented as means ± SEM. N = 3 in each group. *p< 0.05, **p< 0.01.

Article Snippet: The primary antibodies used were anti-Collagen I (1:1000; Zenbio), anti-Collagen Ш (1:1000; Huabio), TGF-β1 (1:1000; Zenbio), α-SMA (1:1000; Zenbio), anti-TLR4 (1:1000; Proteintech), anti-NF-κB P65 (1:1000; Zenbio), anti-P-P65 (1:500; Zenbio), anti-IкBα (1:1000; Zenbio), and anti-p-IкBα (1:500; Zenbio), anti-NLRC4 (1:1000; AiFang), anti-Caspase-1 (1:1000; Zenbio), anti-ASC (1:1000; Zenbio), anti-IL-1β (1:500; Huabio), anti-GAPDH (1:50000; Proteintech).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: PLOS ONE

Article Title: MICT ameliorates hypertensive nephropathy by inhibiting TLR4/NF-κB pathway and down-regulating NLRC4 inflammasome

doi: 10.1371/journal.pone.0306137

Figure Lengend Snippet: (B) CCK-8 assay was used to determine the cell viability. We found that 10 μM Ang II significantly reduced the cell viability of HK-2 cells. We selected 10 μM Ang II for inducing HN model in vitro based on the results of CCK-8. HK-2 cells were treated with 1, 3, 5 μM TAK-242 for 2 hours prior to 10 μM Ang II for 24 hours. (A) Western blot was used to detect the protein levels of (C) TLR4, (D) p-p65 and p65, (E) p-IκBα and IκBα. Data are presented as means ± SEM. N = 3 in each group. *p< 0.05, **p< 0.01, ***p< 0.001.

Article Snippet: The primary antibodies used were anti-Collagen I (1:1000; Zenbio), anti-Collagen Ш (1:1000; Huabio), TGF-β1 (1:1000; Zenbio), α-SMA (1:1000; Zenbio), anti-TLR4 (1:1000; Proteintech), anti-NF-κB P65 (1:1000; Zenbio), anti-P-P65 (1:500; Zenbio), anti-IкBα (1:1000; Zenbio), and anti-p-IкBα (1:500; Zenbio), anti-NLRC4 (1:1000; AiFang), anti-Caspase-1 (1:1000; Zenbio), anti-ASC (1:1000; Zenbio), anti-IL-1β (1:500; Huabio), anti-GAPDH (1:50000; Proteintech).

Techniques: CCK-8 Assay, In Vitro, Western Blot

Journal: PLOS ONE

Article Title: MICT ameliorates hypertensive nephropathy by inhibiting TLR4/NF-κB pathway and down-regulating NLRC4 inflammasome

doi: 10.1371/journal.pone.0306137

Figure Lengend Snippet: HK-2 cells were treated with 1, 3, 5 μM TAK-242 for 2 hours prior to 10 μM Ang II for 24 hours. (A, B) Western blot was used to detect the protein levels of (C-F) fibrosis-related molecules and (G-J) NLRC4 inflammasome. Data are presented as means ± SEM. N = 3 in each group. *p< 0.05, **p< 0.01, ***p< 0.001.

Article Snippet: The primary antibodies used were anti-Collagen I (1:1000; Zenbio), anti-Collagen Ш (1:1000; Huabio), TGF-β1 (1:1000; Zenbio), α-SMA (1:1000; Zenbio), anti-TLR4 (1:1000; Proteintech), anti-NF-κB P65 (1:1000; Zenbio), anti-P-P65 (1:500; Zenbio), anti-IкBα (1:1000; Zenbio), and anti-p-IкBα (1:500; Zenbio), anti-NLRC4 (1:1000; AiFang), anti-Caspase-1 (1:1000; Zenbio), anti-ASC (1:1000; Zenbio), anti-IL-1β (1:500; Huabio), anti-GAPDH (1:50000; Proteintech).

Techniques: Western Blot