Journal: Journal of Histochemistry and Cytochemistry

Article Title: The Expression of Toll-like Receptors in Normal Human and Murine Gastrointestinal Organs and the Effect of Microbiome and Cancer

doi: 10.1369/0022155416656154

Figure Lengend Snippet: Used TLR Antibodies With Dilutions, Catalog Numbers, and Manufacturer.

Article Snippet: TLR3 , 1:25 , IMG-315A , Imgenex.

Techniques:

Journal: Journal of Histochemistry and Cytochemistry

Article Title: The Expression of Toll-like Receptors in Normal Human and Murine Gastrointestinal Organs and the Effect of Microbiome and Cancer

doi: 10.1369/0022155416656154

Figure Lengend Snippet: Typical Toll-like receptor (TLR) expression patterns from the human ascending colon. TLRs are expressed throughout the epithelium with a diffuse manner. Paired figures from ascending colon organ donor (left) and tumor-adjacent normal epithelium (right) are presented: (A) TLR1, (B) TLR2, (C) TLR3, (D) TLR4, (E) TLR5, (F) TLR6, (G) TLR7, (H) TLR8, and (I) TLR9. 20× magnification was used and 50-µm scale bar is in panel I.

Article Snippet: TLR3 , 1:25 , IMG-315A , Imgenex.

Techniques: Expressing

Journal: Journal of Histochemistry and Cytochemistry

Article Title: The Expression of Toll-like Receptors in Normal Human and Murine Gastrointestinal Organs and the Effect of Microbiome and Cancer

doi: 10.1369/0022155416656154

Figure Lengend Snippet: TLR1 to TLR9 Protein Expression Detected With Immunohistochemistry From Different Anatomic Segments of the Alimentary Tract.

Article Snippet: TLR3 , 1:25 , IMG-315A , Imgenex.

Techniques: Expressing, Immunohistochemistry

Journal: Journal of Histochemistry and Cytochemistry

Article Title: The Expression of Toll-like Receptors in Normal Human and Murine Gastrointestinal Organs and the Effect of Microbiome and Cancer

doi: 10.1369/0022155416656154

Figure Lengend Snippet: Typical Toll-like receptor (TLR) expression patterns from the conventional and germ-free mouse small intestines. TLRs are expressed throughout the epithelium with a diffuse manner. Paired figures from small intestine conventional (left) and germ-free mice (right) are presented: (A) TLR1, (B) TLR2, (C) TLR3, (D) TLR4, (E) TLR5, (F) TLR6, (G) TLR7, (H) TLR8, and (I) TLR9. 10× magnification was used and 50-µm scale bar is in panel I.

Article Snippet: TLR3 , 1:25 , IMG-315A , Imgenex.

Techniques: Expressing

Journal: Immunobiology

Article Title: Scavenger receptor SREC-I promotes double stranded RNA-mediated TLR3 activation in human monocytes

doi: 10.1016/j.imbio.2014.12.011

Figure Lengend Snippet: A and B, TLR3 colocalized with SREC-I in HEK 293-TLR3-SREC-I overexpressing cells. HEK293 cells were transfected with FLAG-SREC-I and TLR3-CFP for 22 hours. Cells were then incubated without (A) or with (B) Poly I:C (10ug/ml) on ice for 30 min. Cells (A) and (B) were then fixed with 4% para formaldehyde and permeabilized with 0.1% Triton X100 (A) or not (B). Cells were then stained for FLAG with anti-FLAG M2 antibody (green) C, HEK293 cells were transfected with FLAG-SREC-I and TLR3-CFP for 22 hr. Cells were then incubated Poly I:C (10ug/ml) on ice for 30 min followed by incubation with warm media at 37°C for 20 min. Cells were then fixed with 4% PFA and permeabilized with 0.1% Triton X100. Cells were stained with anti-FLAG antibody (green). D, HEK293 cells were transfected with FLAG-SREC-1 or TLR3-CFP for 18 hours. Cells were incubated with Poly I:C (10μg/ml) for 1 hour and then fixed and stained with anti-FLAG M2 ab (green). E, Differentiated THP1 cells were transfected with FLAG-SREC-I and TLR3-CFP for 22 hr. Cells were then incubated with Poly I:C (10μg/ml) for 1 hr and then fixed and permeabilized as in D. Cells were stained for SREC-I (green), with anti FLAG ab, LAMP1 with anti LAMP1 (red). TLR3-CFP is in blue. Experiments were repeated twice with reproducible findings. Scale bar 2 micron.

Article Snippet: Both human and mice SREC-I (siRNA) and TLR3 (siRNA) constructs were purchased from Santa Cruz Biotech.

Techniques: Transfection, Incubation, Staining

Journal: Immunobiology

Article Title: Scavenger receptor SREC-I promotes double stranded RNA-mediated TLR3 activation in human monocytes

doi: 10.1016/j.imbio.2014.12.011

Figure Lengend Snippet: HEK293 cells stably expressing TLR3 was transfected with SREC-I-GFP for 22 hr. Cells were then treated with or without 10ug/ml Poly I:C for 30 min. Cell lysates were collected and SREC-I-GFP was immunoprecipitated using anti-GFP antibody and anti GFP ab and then the precipitated complexes were subjected to SDS-PAGE analysis followed by blotting for TLR3 using anti-TLR3 antibody. Experiments were carried out twice, reproducibly.

Article Snippet: Both human and mice SREC-I (siRNA) and TLR3 (siRNA) constructs were purchased from Santa Cruz Biotech.

Techniques: Stable Transfection, Expressing, Transfection, Immunoprecipitation, SDS Page

Journal: Immunobiology

Article Title: Scavenger receptor SREC-I promotes double stranded RNA-mediated TLR3 activation in human monocytes

doi: 10.1016/j.imbio.2014.12.011

Figure Lengend Snippet: A, B and C, HEK293-TLR3 cells were transfected with SREC-I or untransfected HEK293-TLR3 cells were then incubated with or without Poly I:C (10μg/ml) for 2 hr. Cell lysates were collected and then subjected to SDS-PAGE and western blotting with appropriate antibodies. D, Bone marrow derived macrophages (BMDM) were transfected with siRNA of SREC-I for 72 hr. Cells were then incubated with Poly I:C (or not) as in A. Cell were lysed and equal amount of protein were subjected to SDS-PAGE and western blotting using anti phospho-p65 antibody and anti p65 antibody. E, BMDM cells were treated as in D and then cell lysates were subjected to SDS-PAGE and western blotting using antibodies shown in E. F, HEK293-TLR3 cells were transfected with SREC-I or not and also NFkβ-SEAP/CMV-SEAP constructs. Cells were incubated with Poly I:C (10μg/ml)/ODN2395 (10μg/ml), a non TLR3 ligand. NFkβ activity was measured as instructed by NFkβ-SEAporter assay kit. Similar results were observed in two separate experiments.

Article Snippet: Both human and mice SREC-I (siRNA) and TLR3 (siRNA) constructs were purchased from Santa Cruz Biotech.

Techniques: Transfection, Incubation, SDS Page, Western Blot, Derivative Assay, Construct, Activity Assay

Journal: Immunobiology

Article Title: Scavenger receptor SREC-I promotes double stranded RNA-mediated TLR3 activation in human monocytes

doi: 10.1016/j.imbio.2014.12.011

Figure Lengend Snippet: A, THP1 cells were transfected with TLR3 or TLR3 and SREC-I expression plasmids for 22 hr. Cells were incubated with 10ug Poly I:C for 12 hr and then assayed for cytokine production using a human cytokine multianylate ELISA array kit according to manufacturer’s protocol. B, THP1 cells treated as in (A) and then assayed for chemokines using multianylate ELISA array kit according to Manufacturer’s protocol. Data represent the mean of two independent experiments.

Article Snippet: Both human and mice SREC-I (siRNA) and TLR3 (siRNA) constructs were purchased from Santa Cruz Biotech.

Techniques: Transfection, Expressing, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Immunobiology

Article Title: Scavenger receptor SREC-I promotes double stranded RNA-mediated TLR3 activation in human monocytes

doi: 10.1016/j.imbio.2014.12.011

Figure Lengend Snippet: A, THP1 cells were transfected with TLR3/SREC-I or TLR3 and SREC-I expression plasmids for 22 hr. Cells were incubated with Poly I:C (10ug/ml) for 12 hr and then assayed for IL-8 release. B, THP1 cells were transfected with SREC-I and TLR3 for 22 hr and then incubated with the ERK inhibitor (PD98059), p38 inhibitor (SB203580) or the JNK inhibitor (JNK inhibitor II) (10uM) for 1 hr right before incubation with incubation with Poly I:C for 12 hr. IL-8 secretion was assayed according to manufacturer’s instructions. C, THP1 cells were treated as in A and then IL-6 release was assayed according to manufacturer’s instruction. D, THP1 cells were treated as in B and IL-6 release was assayed. E, THP1 cells were treated with PMA (5–10ng/ml) for 72 hours. Cells were then transfected with siRNA SREC-I or siRNA TLR3. Cells were incubated with Poly I:C for 12 hr and then Ifnβ release was measured according to manufacturer’s protocol. F. BMDM cells were transfected with siRNA SREC-I/TLR3 or not. Wild type or TLR3 knocked down (siRNA) cells were also treated with 1–5 ng/ml of LPS for 12hrs. Cells were then incubated with Poly I:C for 12 hr. Secreted IL-6 in media was measured according to manufacturer’s protocol. Experiment was repeated twice. Data shown are the mean ±SD of results from those two experiments.

Article Snippet: Both human and mice SREC-I (siRNA) and TLR3 (siRNA) constructs were purchased from Santa Cruz Biotech.

Techniques: Transfection, Expressing, Incubation

Journal: Immunobiology

Article Title: Scavenger receptor SREC-I promotes double stranded RNA-mediated TLR3 activation in human monocytes

doi: 10.1016/j.imbio.2014.12.011

Figure Lengend Snippet: A, THP1-TLR3 cells were transfected with or without SREC-I for 22 hr and then incubated with 10ug Poly I:C for indicated time. IL-8 release from cells was then assayed. B, THP1-TLR3 cells were transfected as in A and then incubated with or without Bafilomycin A (0.2 uM for 20 min) or PP2 (10uM for 12 hr). Cells were incubated with Poly I:C for 12 hr and then IL-8 release from THP1 cells were assayed. C, Differentiated THP1 cells were transfected with siRNA SREC-I for 72 hr and then cells were treated with Poly I:C (10μg/ml) for 2 hr. Cells were then lysed and equal amount of protein in lysate were subjected to SDS-PAGE and western blotting using appropriate antibodies. Experiments were carried out twice reproducibly.

Article Snippet: Both human and mice SREC-I (siRNA) and TLR3 (siRNA) constructs were purchased from Santa Cruz Biotech.

Techniques: Transfection, Incubation, SDS Page, Western Blot

Journal: Immunobiology

Article Title: Scavenger receptor SREC-I promotes double stranded RNA-mediated TLR3 activation in human monocytes

doi: 10.1016/j.imbio.2014.12.011

Figure Lengend Snippet: Double stranded RNA species Poly I:C interacts with macrophages leading to recruitment of SREC-I and TLR3 to the cell surface where they form a membrane complex that interacts with c-Src. The latter kinase then regulates endocytosis of the SREC-I/TLR3/Poly I:C complexes in endosomes. The complex finally resides in endosomes with low intravesicular pH, marked with lysosomal protein LAMP1. TLR3 is able to signal from such complexes and launches NFkB, MAPK and IRF3 signaling. (IRF3 is activated by the kinase TBK1). Activated NFkB, AP-1 and C/EBPβ are known to interact with the IL-8 gene while activated IRF3 leads to synthesis of Ifn-β.

Article Snippet: Both human and mice SREC-I (siRNA) and TLR3 (siRNA) constructs were purchased from Santa Cruz Biotech.

Techniques: Membrane

Journal: Immunity

Article Title: Autoimmune Th17 Cells Induced Synovial Stromal and Innate Lymphoid Cell Secretion of the Cytokine GM-CSF to Initiate and Augment Autoimmune Arthritis

doi: 10.1016/j.immuni.2018.04.009

Figure Lengend Snippet:

Article Snippet: TaqMan Gene Expression ( Tlr3 ) , ABI , Mm01207404_m1.

Techniques: Control, Recombinant, Staining, Enzyme-linked Immunosorbent Assay, Gene Expression, Software, FCAP Assay

Journal: Journal of Neurovirology

Article Title: Expression of miR-155 associated with Toll-like receptors 3, 7, and 9 transcription in the olfactory bulbs of cattle naturally infected with BHV5

doi: 10.1007/s13365-017-0564-6

Figure Lengend Snippet: a Detection of viral DNA amplification in ORNs, GL, and ML from olfactory bulbs suspensions collected from positive and negative animals. b Transcription of TLR3 , TLR7 , TLR9 , and miR-155 by qPCR in ORNs, GL, and ML from the same tissue samples. The data is a media (X) plus standard deviation (s.d) of positive and negative samples analyzed

Article Snippet: Both TaqManTM FAM-MGB primers and probes: TLR3 Bt03210267_m1 (cat # 4351372), TLR7 Bt03225261_s1 (cat # 4351373), and TLR9 Bt03224812_m1 (cat # 4331182).

Techniques: DNA Amplification, Standard Deviation