Journal: The FASEB Journal

Article Title: Mycobacterium tuberculosis induces connective tissue growth factor expression through the TLR2-JNK–AP-1 pathway in human lung fibroblasts

doi: 10.1096/fj.201900487R

Figure Lengend Snippet: TLR2 mediated M.tb-induced CTGF expression in human lung fibroblasts. A) WI-38 cells were pretreated with 30 μg/ml anti-TLR2 antibody and anti-TLR4 antibody for 30 min followed by treatment with 10 μg/ml M.tb for another 1 h. Cells were then immunodetected with CTGF- or α-tubulin–specific antibodies. B) NHLFs were pretreated with 30 μg/ml anti-TLR2 antibody and anti-TLR4 antibody for 30 min followed by treatment with 10 μg/ml M.tb for another 1 h. Cells were then immunodetected with CTGF- or α-tubulin–specific antibodies. Data are expressed as means ± sem from 3 experiments. *P < 0.05 compared with M.tb treatment.

Article Snippet: Antibodies specific for TLR2 and TLR4, rabbit polyclonal IgG, protein A/G beads, c-Jun phosphorylated at Ser63, CTGF, fibronectin, anti-rabbit and anti-goat IgG-conjugated horseradish peroxidase, anti–c-Fos, anti-mouse, and c-Jun antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing

Journal: The FASEB Journal

Article Title: Mycobacterium tuberculosis induces connective tissue growth factor expression through the TLR2-JNK–AP-1 pathway in human lung fibroblasts

doi: 10.1096/fj.201900487R

Figure Lengend Snippet: Diagrammatic sketch summary of the signaling pathway of M.tb-induced CTGF expression. M.tb induced the activation of the TLR2-JNK–c-Jun–AP-1 cascade, which resulted in an increased CTGF expression and ECM production in human lung fibroblasts.

Article Snippet: Antibodies specific for TLR2 and TLR4, rabbit polyclonal IgG, protein A/G beads, c-Jun phosphorylated at Ser63, CTGF, fibronectin, anti-rabbit and anti-goat IgG-conjugated horseradish peroxidase, anti–c-Fos, anti-mouse, and c-Jun antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Activation Assay

Journal: Pathogens

Article Title: The Immunological Changes in the Skin of BALC/c Mice with Disseminated Acanthamoebiasis

doi: 10.3390/pathogens12050631

Figure Lengend Snippet: The TLR2 gene expression in the skin of control and amoeba-infected mice at 8, 16 and 24 dpi, (A, amoeba-infected host with normal immunity; AS, amoeba-infected host with reduced immunity induced by MPS, methylprednisolone sodium succinate; C, control host with normal immunity; CS, control host with reduced immunity induced by MPS). In the graph arithmetic mean ± standard deviation was shown; p < 0.05.

Article Snippet: Immunohistochemistry was performed using specific primary rabbit polyclonal antibodies against TLR2 and TLR4 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; cat. no. sc-10739 and sc-30002, respectively) in a final 1:500 dilution, after checking and standardized other recommended by manufacturer concentrations from 1:50 to 1:500.

Techniques: Expressing, Infection, Standard Deviation

Journal: Pathogens

Article Title: The Immunological Changes in the Skin of BALC/c Mice with Disseminated Acanthamoebiasis

doi: 10.3390/pathogens12050631

Figure Lengend Snippet: Representative microphotography that shows immunoexpression of TLR2 ( A – H ) and TLR4 ( I – P ) in skin of immunocompetent and immunosuppressed uninfected mice (0 dpi) and at 8, 16, and 24 days after Acanthamoeba spp. infection (dpi). Immunohistochemical reaction with diaminobenzidine (DAB) as a chromogen. Objective magnification 40×. Red arrows mark the area of connective tissue of dermis where immunoexpression of Toll-like receptors 2 (TLR2) and 4 (TLR4) was visible as brown pigmentation; white arrows mark the keratinocytes of epidermis where immunoexpression of TLR2 was visible as brown pigmentation. Abbreviation: C—uninfected immunocompetent mice (n = 5), CS—uninfected immunosuppressed mice (n = 5), A—immunocompetent Acanthamoeba spp. infected mice (n = 5), AS—immunosuppressed Acanthamoeba spp. infected mice (n = 5).

Article Snippet: Immunohistochemistry was performed using specific primary rabbit polyclonal antibodies against TLR2 and TLR4 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; cat. no. sc-10739 and sc-30002, respectively) in a final 1:500 dilution, after checking and standardized other recommended by manufacturer concentrations from 1:50 to 1:500.

Techniques: Infection, Immunohistochemical staining

Journal: Parasitology Research

Article Title: Toll-like receptors in the brain of mice following infection with Acanthamoeba spp.

doi: 10.1007/s00436-016-5217-9

Figure Lengend Snippet: Expression of TLR2 gene at the mRNA level in brains isolated from uninfected and Acanthamoeba -infected mice from patient with Acanthamoeba keratitis (strain Ac55; a ) and Malta Lake (strain Ac43; b ). Brains were dissected from mice at 2, 4, 16, and 30 dpi. Expression level of TLR2 gene was determined by Q-PCR relative quantification analysis evaluated using a calibrator (cDNA mix from all samples). The quantify of TLR2 transcript in each sample was standardized to the amount of PBGD cDNA as the internal control. The amounts of TLR2 mRNA are expressed as the multiplicity of these cDNA concentrations in the calibrator. Each sample was determined in triplicate. Data represent mean ± SD and are representative of groups of six animals in an experiment. * P < 0.05, compared with the control value derived from uninfected mice (Student’s t test)

Article Snippet: Immunohistochemistry was performed using specific primary rabbit polyclonal antibodies against TLR2 and TLR4 (Santa Cruz Biotechnology, Inc., cat. no. sc-10739 and sc-30002) in a final 1:500 dilution.

Techniques: Expressing, Isolation, Infection, Quantitative Proteomics, Control, Derivative Assay

Journal: Parasitology Research

Article Title: Toll-like receptors in the brain of mice following infection with Acanthamoeba spp.

doi: 10.1007/s00436-016-5217-9

Figure Lengend Snippet: Immunoexpression of Toll-like receptor 2 (TLR2) ( a , c , d , g , h , k , l , o , p ) and Toll-like receptor 4 (TLR4) ( b , e , f , i , j , m , n , q , r ) within neocortex of control ( a , b ) and mice infected with Acanthamoeba spp. isolated from patient with Acanthamoeba keratitis strain Ac55 ( c , e , g , i , k , m , o , q ) and from Malta Lake strain Ac43 ( d , f , h , j , l , n , p , r ) in 2, 4, 16, and 30 dpi. Exemplary immunopositive cells: neurons— black arrows ; glial cells— blue arrows ; endothelial cells of neural capillaries— red arrows . Intensity of IHC reaction was highest in the 2-dpi group and decreased during the period of infection. Objective magnification ×40

Article Snippet: Immunohistochemistry was performed using specific primary rabbit polyclonal antibodies against TLR2 and TLR4 (Santa Cruz Biotechnology, Inc., cat. no. sc-10739 and sc-30002) in a final 1:500 dilution.

Techniques: Control, Infection, Isolation

Journal: Parasitology Research

Article Title: Toll-like receptors in the brain of mice following infection with Acanthamoeba spp.

doi: 10.1007/s00436-016-5217-9

Figure Lengend Snippet: Immunoexpression of Toll-like receptor 2 (TLR2) ( a , b , e , f ) and Toll-like receptor 4 (TLR4) ( c , d , g , h ) within choroid plexus of mice infected with Acanthamoeba spp. isolated from patient with Acanthamoeba keratitis (strain Ac55) ( a , c , e , g ) and from Malta Lake (strain Ac43) ( b , d , f , h ) at 2 and 30 dpi. Choroidal ependymocytes— black arrows ; interstitial cells (possibly dendritic cells)— blue arrows . Intensity of IHC reaction was the highest in 2 dpi groups and very low in 30 dpi groups. Objective magnification ×40

Article Snippet: Immunohistochemistry was performed using specific primary rabbit polyclonal antibodies against TLR2 and TLR4 (Santa Cruz Biotechnology, Inc., cat. no. sc-10739 and sc-30002) in a final 1:500 dilution.

Techniques: Infection, Isolation