tlr5  (Proteintech)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The effect of Massa Medicata Fermentata on the cytokine secretion of colonic mucosa and visceral sensitivity in rats with IBS-D

doi: 10.3389/fcimb.2025.1616796

Figure Lengend Snippet: Intestinal permeability and expression of flagellin and TLR5 protein in the lamina propria of the mucosa of rats in various groups. FITC-Dextran was used to detect the intestinal permeability of rats and laser confocal microscopy was employed to observe the localization of flagellin and TLR5 proteins of colon three repeated tests. (Adopt Tukey statistical method. Refer to the annotations in the reference image for grouping identification.)

Article Snippet: MMF (China Resources Sanjiu Co., Ltd); glacial acetic acid (10000208 Sinopharm, Inc.); FITC-Dextran (Sigma, Inc.); WB Antibodies: TLR5 Antibody (rabbit source, Proteintech, Inc.), TRIF Antibody (rabbit source, Affinity, Inc.); P-ERK1/2 Antibody (rabbit source, abcam, Inc.); immunofluorescent antibodies: immunofluorescent primary antibody: TLR5 (rabbit source, Three Eagles, Inc.), immunofluorescent primary antibody: Flic (mouse source, Yiqiao, Inc.), and Immunofluorescence secondary antibody: goat anti-rabbit IgGH&L (Cy3) (abcam, Inc.), immunofluorescence secondary antibody: goat anti-mouse IgGH&L (FITC) (abcam, Inc.); Immunohistochemistry antibody: immunohistochemistry primary antibody: TLR5 (rabbit source, proteintech, Inc.), immunohistochemistry primary antibody: TRIF (rabbit source, Affinity, Inc.), the Immunohistochemical primary antibody: p-ERK1/2 (rabbit source abcam), immunohistochemical secondary antibody: HRP-labeled goat anti-rabbit IgG (Biyun Tian, Inc.); Percoll lymphocyte isolate P8370 (Solarbio, Inc.); LPDCs magnetic bead sorting kit (OX62) (MiltenyiBiotec, Inc.) Anti-RatCD103, PE (Bioscience, Inc.); Rat spleen lymphocyte isolate LTS1083PK-200 (TBD, Inc.); CD4 Magnetic Bead Sorting Kit (MiltenyiBiotec, Inc.); CD4MonoclonalAntibody;CCK-8 Kit C0039 (Biotronix, Inc.); Rat Interleukin 12 (IL-12) ELISA Kit, Rat Interferon Gamma (IFN-γ) ELISA Kit, Rat Interleukin 4 (IL-4) ELISA Kit, Rat Interleukin 9 (IL-9) kit, Rat Interleukin 6 (IL-6) ELISA kit, Rat Interleukin 17A (IL-17A) ELISA kit, Rat Transforming Growth Factor β (TGF-β) ELISA kit (Jiangsu Jingmei, Inc.).

Techniques: Permeability, Expressing, Confocal Microscopy

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The effect of Massa Medicata Fermentata on the cytokine secretion of colonic mucosa and visceral sensitivity in rats with IBS-D

doi: 10.3389/fcimb.2025.1616796

Figure Lengend Snippet: Expression of TLR5, TRIF and p-EPK1/2 in LPDCs of rats in each group. The mRNA expression of TLR5, TRIF, and p-EPK1/2 in LPDCs was detected by qPCR. The expression of TLR5, TRIF, and p-EPK1/2 proteins in LPDCs of rats in each group was detected by WB three repeated tests. (Adopt Tukey statistical method. Refer to the annotations in the reference image for grouping identification.)

Article Snippet: MMF (China Resources Sanjiu Co., Ltd); glacial acetic acid (10000208 Sinopharm, Inc.); FITC-Dextran (Sigma, Inc.); WB Antibodies: TLR5 Antibody (rabbit source, Proteintech, Inc.), TRIF Antibody (rabbit source, Affinity, Inc.); P-ERK1/2 Antibody (rabbit source, abcam, Inc.); immunofluorescent antibodies: immunofluorescent primary antibody: TLR5 (rabbit source, Three Eagles, Inc.), immunofluorescent primary antibody: Flic (mouse source, Yiqiao, Inc.), and Immunofluorescence secondary antibody: goat anti-rabbit IgGH&L (Cy3) (abcam, Inc.), immunofluorescence secondary antibody: goat anti-mouse IgGH&L (FITC) (abcam, Inc.); Immunohistochemistry antibody: immunohistochemistry primary antibody: TLR5 (rabbit source, proteintech, Inc.), immunohistochemistry primary antibody: TRIF (rabbit source, Affinity, Inc.), the Immunohistochemical primary antibody: p-ERK1/2 (rabbit source abcam), immunohistochemical secondary antibody: HRP-labeled goat anti-rabbit IgG (Biyun Tian, Inc.); Percoll lymphocyte isolate P8370 (Solarbio, Inc.); LPDCs magnetic bead sorting kit (OX62) (MiltenyiBiotec, Inc.) Anti-RatCD103, PE (Bioscience, Inc.); Rat spleen lymphocyte isolate LTS1083PK-200 (TBD, Inc.); CD4 Magnetic Bead Sorting Kit (MiltenyiBiotec, Inc.); CD4MonoclonalAntibody;CCK-8 Kit C0039 (Biotronix, Inc.); Rat Interleukin 12 (IL-12) ELISA Kit, Rat Interferon Gamma (IFN-γ) ELISA Kit, Rat Interleukin 4 (IL-4) ELISA Kit, Rat Interleukin 9 (IL-9) kit, Rat Interleukin 6 (IL-6) ELISA kit, Rat Interleukin 17A (IL-17A) ELISA kit, Rat Transforming Growth Factor β (TGF-β) ELISA kit (Jiangsu Jingmei, Inc.).

Techniques: Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Modulation of the Toll-like Receptor Pathway in Ovine Endometria During Early Pregnancy

doi: 10.3390/ani15070917

Figure Lengend Snippet: Antibodies used for Western blot.

Article Snippet: Mouse anti-TLR5 monoclonal antibody , sc-517439 , Santa Cruz Biotechnology, Santa Cruz, CA, USA , 0.2 μg/mL.

Techniques: Western Blot

Journal: Animals : an Open Access Journal from MDPI

Article Title: Modulation of the Toll-like Receptor Pathway in Ovine Endometria During Early Pregnancy

doi: 10.3390/ani15070917

Figure Lengend Snippet: Relative expression values of ( A ) TLR2 , ( B ) TLR3 , ( C ) TLR4 , ( D ) TLR5 , ( E ) IRAK1 , ( F ) TRAF6 , and ( G ) MYD88 mRNA in the endometrium. Significant differences ( p < 0.05) are indicated by different letters (a, b, c, and d).

Article Snippet: Mouse anti-TLR5 monoclonal antibody , sc-517439 , Santa Cruz Biotechnology, Santa Cruz, CA, USA , 0.2 μg/mL.

Techniques: Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Modulation of the Toll-like Receptor Pathway in Ovine Endometria During Early Pregnancy

doi: 10.3390/ani15070917

Figure Lengend Snippet: Expression of TLR2, TLR3, TLR4, TLR5, IRAK1, TRAF6, and MyD88 proteins in the endometrium. Significant differences ( p < 0.05) are indicated by different letters (a, b, c, and d).

Article Snippet: Mouse anti-TLR5 monoclonal antibody , sc-517439 , Santa Cruz Biotechnology, Santa Cruz, CA, USA , 0.2 μg/mL.

Techniques: Expressing

Journal: iScience

Article Title: Stenotrophomonas maltophilia provokes NEU1-mediated release of a flagellin-binding decoy receptor that protects against lethal infection

doi: 10.1016/j.isci.2024.110866

Figure Lengend Snippet: Sm flagellin binds to the MUC1-ED (A) HEK293T cells were transfected with the pcDNA3.1 empty vector, or plasmids encoding for TLR5 (pTLR5) or MUC1 (pMUC1), the cells were lysed and the lysates processed for TLR5 (lanes 1, 2) or MUC1 (lanes 3, 4) immunoblotting. To control for protein loading and transfer, the blots were stripped and reprobed (IB∗) for β-tubulin. (B) HEK293T cells overexpressing TLR5 or MUC1, or empty vector-transfected cells, were incubated with Sm and adherent bacteria quantified. Vertical bars represent mean ± SEM CFUs/well. (C) A549 cells were transfected with pcDNA3.1 or pMUC1 and binding of increasing concentrations of the Alexa Fluor 594-labeled Sm FliC1,2,3 flagellin mixture determined. Data points represent mean ± SEM bound flagellin in molecules/EC. (D) Scatchard analysis of binding data in (C). The linear regression equation and R 2 value are indicated adjacent to each line. (E and F) HEK293T cells were infected with Ad-GFP or Ad-MUC1, and A549 cells were transfected with MUC1-targeting or control siRNAs, and both cell types were cultured for 48 h. (E) Ad-GFP- and Ad-MUC1-infected cells (lanes 1, 2) and MUC1 siRNA- and control siRNA-transfected cells (lanes 3, 4) were lysed and the lysates processed for MUC1 immunoblotting. (F) Infected or transfected cells were incubated with Sm and cell-bound bacteria quantified. Vertical bars represent mean ± SEM Sm adhesion in CFUs/well. (G) A549 cells were transfected with MUC1-targeting or control siRNAs, cultured for 48 h, and binding of increasing concentrations of the Alexa Fluor 595-labeled Sm FliC1,2,3 flagellin mixture determined. Data points represents mean ± SEM bound flagellin in molecules/EC. (H) Scatchard analysis of binding data in (G). (I–K) A549 cells and A549 cells transfected with MUC1-targeting or control siRNAs were incubated for 48 h and then stimulated for 1 h with 10 ng of the Sm FliC1,2,3 flagellin mixture or PBS. (I) Non-transfected, PBS-treated (lane 1), control siRNA-transfected, flagellin-stimulated (lane 2), and MUC1 siRNA-transfected, flagellin-stimulated (lane 3) cells were lysed, and the lysates processed for pERK1/2 immunoblotting. To control for protein loading and transfer, the blots were stripped and reprobed for total ERK2. The ∼44.0 kDa pERK1 (upper) and ∼42.0 kDa pERK2 (lower) bands, as well as the total ERK2 band, are indicated by arrowheads on the right. (J) Densitometric analysis of pERK1/2 normalized to total ERK2 displayed in (I). Vertical bars represent mean ± SEM normalized pERK1/2 signal. (K) Supernatants from the cells in (I) were processed for IL-8 levels and normalized to total EC protein. Vertical bars represent mean ± SEM normalized IL-8 concentrations in pg/mg EC protein. ∗, increased Sm adhesion (B, F), flagellin binding (C), ERK activation (J), or normalized IL-8 release (K) compared to pcDNA3.1, pTLR5, Ad-GFP, control siRNA, or PBS controls at p < 0.05. ∗∗, decreased Sm adhesion (F), flagellin binding (G), ERK activation (J), or normalized IL-8 release (K) compared to the siRNA controls at p < 0.05. n.s., not significant. Statistical comparisons were made using the Student’s t test. The results are representative of 3 or 6 independent experiments.

Article Snippet: Mouse anti-human TLR5 , Santa Cruz Biotechnology , Cat# sc-52963; RRID: AB_793184.

Techniques: Transfection, Plasmid Preparation, Western Blot, Control, Incubation, Bacteria, Binding Assay, Labeling, Infection, Cell Culture, Activation Assay

Journal: iScience

Article Title: Stenotrophomonas maltophilia provokes NEU1-mediated release of a flagellin-binding decoy receptor that protects against lethal infection

doi: 10.1016/j.isci.2024.110866

Figure Lengend Snippet:

Article Snippet: Mouse anti-human TLR5 , Santa Cruz Biotechnology , Cat# sc-52963; RRID: AB_793184.

Techniques: Virus, Recombinant, Control, Software, Plasmid Preparation, Enzyme-linked Immunosorbent Assay