Journal: Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association

Article Title: Expression of Toll-like receptor 9 in renal podocytes in childhood-onset active and inactive lupus nephritis.

doi: 10.1093/ndt/gfq058

Figure Lengend Snippet: Fig. 1. Immunohistochemistry for TLR9 and synaptopodin in active LN and normal control. In active LN (B, C, arrow, arrowhead; Patient 2) but not in the normal control (A), some glomerular cells strongly expressed TLR9. Some tubular cells were also weakly stained in active LN (asterisk in B). Most of the TLR9-positive cells localized adherent to outside of glomerular capillaries (B and C arrow). In the normal control kidney, synaptopodin was strongly and evenly stained (D) whereas weakly and irregularly stained in active LN (E; Patient 2).

Article Snippet: The tissue sections were stained with monoclonal anti-synaptopodin antibody (G1D4, Progen, Heidelberg, Germany) and monoclonal anti-TLR9 antibody (26C593.2, Imgenex, San Diego, CA, USA).

Techniques: Immunohistochemistry, Control, Staining

Journal: Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association

Article Title: Expression of Toll-like receptor 9 in renal podocytes in childhood-onset active and inactive lupus nephritis.

doi: 10.1093/ndt/gfq058

Figure Lengend Snippet: Fig. 2. Double immunofluorescence staining for TLR9 and synaptopodin in LN and normal control. The TLR9-positive cells stained positively in active LN (D, J; Patient 3), but negatively in remission of LN (G; Patient 3) and a normal control (A). Synaptopodin stained weakly and irregularly in active LN (E, K; Patient 3) compared to remission of LN (H; Patient 3) and a normal control (B). The glomerular cells expressing TLR9 are specifically consistent with synaptopodin-positive cells, namely podocytes (F, L).

Article Snippet: The tissue sections were stained with monoclonal anti-synaptopodin antibody (G1D4, Progen, Heidelberg, Germany) and monoclonal anti-TLR9 antibody (26C593.2, Imgenex, San Diego, CA, USA).

Techniques: Double Immunofluorescence Staining, Control, Staining, Expressing

Journal: PLoS ONE

Article Title: Recruitment of Cbl-b to B Cell Antigen Receptor Couples Antigen Recognition to Toll-Like Receptor 9 Activation in Late Endosomes

doi: 10.1371/journal.pone.0089792

Figure Lengend Snippet: (A) Representative confocal microscopic images of splenocytes from mice with indicated genotypes. For experiments, cells were stimulated through the BCR (green) for 30 minutes then fixed and stained for TLR9 (blue) and Lamp-1 (red)(n = 3). (B) Quantitation of experiments shown in (A) for fraction of cells demonstrating significant co-localization of TLR9 with Lamp-1 (n = 3, 30 cells/exp) (Cbl-b −/− vs. Cblb C373A or WT, p<0.001). (C) Quantitation of fractions of cells in (A) demonstrating significant co-localization between BCR and TLR9 (n = 3, 30 cells/exp). (D) In vitro assay of T-bet induction in response to ODN 1826 or control ODN targeted through the BCR (n = 3, p<0.01).

Article Snippet: Antibodies used for visualization were Lamp-1 (1D4B) (BD Biosciences), TLR9 (26C593.2) (Imgenex), Cathepsin L (CPLH-3G10) and Cbl-b (H-121 and H-454) (Santa Cruz Biotechnology).

Techniques: Staining, Quantitation Assay, In Vitro, Control

Journal: Immunology

Article Title: Identification of two subpopulations of purified human blood B cells, CD27- CD23+ and CD27high CD80+, that strongly express cell surface Toll-like receptor 9 and secrete high levels of interleukin-6.

doi: 10.1111/j.1365-2567.2008.02844.x

Figure Lengend Snippet: Figure 1. Cell-surface Toll-like receptor 9 (TLR9) expression on B cells sorted from peripheral blood mononuclear cells (PBMCs). (a) Western blot analysis of TLR9 protein expressed in human PBMC lysate (lane 1) and positive control (lane 2). The estimated molecular weight of TLR9 is 120 000; b-actin was used as the load- ing control. The Western blot was performed three times with simi- lar results. (b) TLR9 was detected by antibody labelling and flow cytometry analysis after gating for CD19+. (c) Summary of flow cytometry analysis of TLR9 expression by CD19+ B cells. The mean percentage of CD19+ B cells that were positive and negative for TLR9 expression is shown (mean ± SD from 10 independent experi- ments). (d) Detection of cell surface and intracellular TLR9 expres- sion by flow cytometry. Flow cytometry representations of TLR9 expression on B-cell surface and intracellular locations (one repre- sentative experiment out of three). Cells stained with the isotype control antibody provided the background level of TLR9 expression.

Article Snippet: The blots were then incubated with a primary mAb to TLR9 (clone 26C593; Imgenex) for 90 min at room temperature.

Techniques: Expressing, Western Blot, Positive Control, Molecular Weight, Control, Cytometry, Flow Cytometry, Staining

Journal: Immunology

Article Title: Identification of two subpopulations of purified human blood B cells, CD27- CD23+ and CD27high CD80+, that strongly express cell surface Toll-like receptor 9 and secrete high levels of interleukin-6.

doi: 10.1111/j.1365-2567.2008.02844.x

Figure Lengend Snippet: Figure 3. Detection of interleukin-6 (IL-6) in the culture superna- tants of the indicated B-cell subsets stimulated with oligodeoxydi- nucleotide (ODN; CpG 2006, CpG 2006-G5) or control ODN (CpG 2006c) for 48 hr. (a) CD19+ CD27) CD23+ TLR9+ and CD19+

Article Snippet: The blots were then incubated with a primary mAb to TLR9 (clone 26C593; Imgenex) for 90 min at room temperature.

Techniques: Control

Journal: Immunologic research

Article Title: FGL2 knockdown improves heart function through regulation of TLR9 signaling in the experimental autoimmune myocarditis rats

doi: 10.1007/s12026-017-8965-4

Figure Lengend Snippet: Western blotting analysis for TLR9 and Fgl2 protein expression in rat myocardial tissue. a TLR9 and Fgl2 protein expression stripe in each group. TLR9 protein expression in myocardial tissue of the EAM group was significantly higher than that of the control group on day 21 after initial immunization. TLR9 protein expression was significantly lower in the RNAi group compared with the EAM group, and also significantly higher in the RNAi group compared with the NC group. Fgl2 protein expression in myocardial tissue of the EAM group was significantly higher than that in the NC group on day 21 after initial immunization. Fgl2 protein expression was significantly lower compared with those in EAM and GFP groups. b β-Actin as control; the TLR9/β-actin and Fgl2/β-actin ratio of NC was considered as 100%. *P < 0.05 vs NC, group, #P < 0.05 vs EAM group

Article Snippet: Recombinant virus’ plasmid and the carriers of auxiliary packaging original plasmids were purchased from Shanghai Jikai Gene Chemical Co.. PCR re-agents were as follows: TRIzol kits were purchased from Invitrogen, retrovirus RNA kits from Promega, and RNase free from Axygen; PCR primer sequences were designed and synthesized by Shanghai Promega Biological company; P1250 Protein Extraction Kit I was purchased from Applygen Technologies Inc.; rat anti-Fgl2 antibody was purchased from Sigma; rat anti-TLR9 antibody was purchased from Novus Biologicals; cardiac myosin of pigs’ heart was purchased from Sigma, complete Freund’s adjuvant CFA was purchased from Sigma; and rat anti-β-actin antibody and goat anti-rat IgG antibody were purchased from ZSGB-BIO.

Techniques: Western Blot, Expressing, Control

Journal: Immunologic research

Article Title: FGL2 knockdown improves heart function through regulation of TLR9 signaling in the experimental autoimmune myocarditis rats

doi: 10.1007/s12026-017-8965-4

Figure Lengend Snippet: Quantitative RT-PCR was used to detect the level of TLR9 and CTLA-4 mRNA. TLR9 and CTLA-4 mRNA expressions in myocardial tissue of the EAM group were significantly higher than in the control group on day 21 after initial immunization. The expression level was significantly higher in the RNAi group compared with the control group but was lower compared with the EAM group. *P < 0.05 vs NC group, #P < 0.05 vs EAM group

Article Snippet: Recombinant virus’ plasmid and the carriers of auxiliary packaging original plasmids were purchased from Shanghai Jikai Gene Chemical Co.. PCR re-agents were as follows: TRIzol kits were purchased from Invitrogen, retrovirus RNA kits from Promega, and RNase free from Axygen; PCR primer sequences were designed and synthesized by Shanghai Promega Biological company; P1250 Protein Extraction Kit I was purchased from Applygen Technologies Inc.; rat anti-Fgl2 antibody was purchased from Sigma; rat anti-TLR9 antibody was purchased from Novus Biologicals; cardiac myosin of pigs’ heart was purchased from Sigma, complete Freund’s adjuvant CFA was purchased from Sigma; and rat anti-β-actin antibody and goat anti-rat IgG antibody were purchased from ZSGB-BIO.

Techniques: Quantitative RT-PCR, Control, Expressing

Journal: BMC Cancer

Article Title: TLR9 expression in glioma tissues correlated to glioma progression and the prognosis of GBM patients

doi: 10.1186/1471-2407-10-415

Figure Lengend Snippet: Expression of TLR9 in glioma cell lines and glioma samples . A . mRNA expression of TLR9 (a) Expression of TLR9 in human U87 glioma cell line(lane1) and human U251 glioma cell line(lane2); (b) Expression of TLR9 in rat glioma cell line C6. (c) Expression of TLR9 in 34 clinical glioma samples (lane1-15, Grade IV gliomas; lane16-28, grade III gliomas;lane 29-34, grade II gliomas). GAPDH was used as positive control. B . Protein expression of TLR9 in human glioma cell lines U87 and U251 with immunofluorescence staining. cytoplasmic localization of TLR9(red fluorescence, Cy3) in U87 and U251 was confirmed by confocal microscopy, nucleus was stained with Hoechst(Blue fluorescence). (a) TLR9 staining in U87 cells. (b) Hoechst staining of nucleus in U87 cells. (c) The combined figure of a and b. (d) negative control, staining of U87 cells with secondary antibody and Hoechst, but without primary anti-TLR9 antibody. (e) TLR9 staining in U251 cells. (f) Hoechst staining of nucleus in U251 cells. (g) The combined figure of e and f. (h) Negative control, staining of U251 cells with secondary antibody and Hoechst, but without primary anti-TLR9 antibody.

Article Snippet: Cells were incubated overnight at 4°C with TLR9 mAb (Img-305A, clone 26C593.2, Imgenex, San Diego, USA, 1:400 dilution), followed by Cy3-labelled anti-mouse secondary antibody (Sigma, USA, 1:400 dilution) for 2 h. The nuclei were counterstained with Hoechst (1:1000 dilutions).

Techniques: Expressing, Positive Control, Immunofluorescence, Staining, Fluorescence, Confocal Microscopy, Negative Control

Journal: BMC Cancer

Article Title: TLR9 expression in glioma tissues correlated to glioma progression and the prognosis of GBM patients

doi: 10.1186/1471-2407-10-415

Figure Lengend Snippet: TLR9 expression in glioma tissues correlated to glioma progression and the prognosis of GBM patients . A . Representative immunohistochemistry staining photos of the glioma tissue arrays. Normal human kidney tissues provided by Department of Surgery were used as positive control. For negative controls, the primary antibody was replaced by normal mouse serum. Cell nuclei were counterstained with haematoxylin. B . Kaplan-Meier analysis showing the PFS(progress free survival) of glioblatoma (GBM) patients in low TLR9 expression group (bold line) and high TLR9 expression group (dotted line). The difference of PFS between the two groups is significant(p = 0.001).

Article Snippet: Cells were incubated overnight at 4°C with TLR9 mAb (Img-305A, clone 26C593.2, Imgenex, San Diego, USA, 1:400 dilution), followed by Cy3-labelled anti-mouse secondary antibody (Sigma, USA, 1:400 dilution) for 2 h. The nuclei were counterstained with Hoechst (1:1000 dilutions).

Techniques: Expressing, Immunohistochemistry, Staining, Positive Control

Journal: BMC Cancer

Article Title: TLR9 expression in glioma tissues correlated to glioma progression and the prognosis of GBM patients

doi: 10.1186/1471-2407-10-415

Figure Lengend Snippet: TLR9 expression is correlated with glioma progression

Article Snippet: Cells were incubated overnight at 4°C with TLR9 mAb (Img-305A, clone 26C593.2, Imgenex, San Diego, USA, 1:400 dilution), followed by Cy3-labelled anti-mouse secondary antibody (Sigma, USA, 1:400 dilution) for 2 h. The nuclei were counterstained with Hoechst (1:1000 dilutions).

Techniques: Expressing

Journal: BMC Cancer

Article Title: TLR9 expression in glioma tissues correlated to glioma progression and the prognosis of GBM patients

doi: 10.1186/1471-2407-10-415

Figure Lengend Snippet: Univariate and multivariate analysis of different prognostic parameters for PFS of GBM patients

Article Snippet: Cells were incubated overnight at 4°C with TLR9 mAb (Img-305A, clone 26C593.2, Imgenex, San Diego, USA, 1:400 dilution), followed by Cy3-labelled anti-mouse secondary antibody (Sigma, USA, 1:400 dilution) for 2 h. The nuclei were counterstained with Hoechst (1:1000 dilutions).

Techniques: Expressing