Journal: Experimental and Therapeutic Medicine

Article Title: Probiotic effect on Helicobacter pylori attachment and inhibition of inflammation in human gastric epithelial cells

doi: 10.3892/etm.2019.7742

Figure Lengend Snippet: TLR4, IκBα and NF-κB p65 expression by GES-1 cells treated with HP-LPS in the absence or presence of live L. bulgaricus . (A) Representative immunoblots of NF-κB p65 compared with lamin B1 in nuclear extracts. (B) Representative immunoblots of TLR4 and IκBα compared with GAPDH in cytoplasm extracts. (C) Quantification of NF-κB p65, (D) IκBα and (E) TLR4 protein levels following different treatments. *P<0.05 vs. HP-LPS 60 min group; # P<0.05 vs. HP-LPS 120 min group. NF-κB, nuclear factor-κB; TLR4, toll-like receptor 4; IκBα, NFKB inhibitor-α; HP, Helicobacter pylori ; LPS, lipopolysaccharide; L, Lactobacillus .

Article Snippet: Membranes were blocked at room temperature with 5% fat-free dried milk in Tris buffered saline containing 0.1% Tween-20 (TBST) for 2 h. Following this, membranes were incubated overnight at 4°C with primary antibodies anti-toll-like receptor 4 (TLR4; cat. no. AF1478-SP; 1:1,000; R&D Systems, Inc.), anti-NF-κB p65 (cat. no. 8242P; 1:1,000; Cell Signaling Technology, Inc.), and anti-NFKB inhibitor-α (IκBα; cat. no. 4814P; 1:1,000; Cell Signaling Technology, Inc.).

Techniques: Expressing, Western Blot

Journal: Cancer Cell International

Article Title: The same and not the same: heterogeneous functional activation of prostate tumor cells by TLR ligation

doi: 10.1186/1475-2867-14-54

Figure Lengend Snippet: Expression of TLRs in human prostate cancer cell lines. A : RT-PCR showing expression of TLR1-10, MyD88 and CD14 transcripts. RT-PCR was performed according to the protocol described in Materials and methods. PBMC served as positive control. Amplification of Beta-actin in the same samples was used as loading control. No amplification controls (NC)(no reverse transcriptase added) were included as negative controls in all PCR programs. B : Western blot analysis of TLR2 and TLR4 expression. Positive sample and loading controls were included as RT-PCR experiments. Lanes in which primary antibodies were substituted by non-immune sera served as negative controls (NC). Similar results were observed in three independent experiments.

Article Snippet: The membranes were blocked with 5% non-fat dry milk and probed with 1:100 dilution of goat anti-human TLR2 (Santa Cruz, USA) or 1 μg/mL goat anti-human TLR4 (R&D, USA) for 2 h. Membranes were consequently incubated with peroxidase-conjugated rabbit anti-goat secondary antibody (Avicenna research Institute, Iran) at a dilution of 1:10000.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Amplification, Western Blot

Journal: Cancer Cell International

Article Title: The same and not the same: heterogeneous functional activation of prostate tumor cells by TLR ligation

doi: 10.1186/1475-2867-14-54

Figure Lengend Snippet: Flow cytometric analysis of TLR2 and TLR4 expression in human prostate cancer cell lines. A : Basal levels of TLR2, TLR4 and CD14 expression was assessed using specific antibodies (white areas) or isotype controls (gray areas). Monocyte gate of PBMC served as positive control. B : Cells were stimulated with a combination of LPS + IL-1 or LTA + IL-1 and expression of TLR2 and TLR4 was evaluated as above. The diagrams are representative of at least three independent experiments. C : Immunofluorescent localization of TLR2 and TL4 in prostate cancer cell lines. MACS-purified human monocytes and HL-60 cell line served as positive control for TLR2 and TLR4 expression, respectively. Scale bare: 50 μm.

Article Snippet: The membranes were blocked with 5% non-fat dry milk and probed with 1:100 dilution of goat anti-human TLR2 (Santa Cruz, USA) or 1 μg/mL goat anti-human TLR4 (R&D, USA) for 2 h. Membranes were consequently incubated with peroxidase-conjugated rabbit anti-goat secondary antibody (Avicenna research Institute, Iran) at a dilution of 1:10000.

Techniques: Expressing, Positive Control, Purification

Journal: Cancer Cell International

Article Title: The same and not the same: heterogeneous functional activation of prostate tumor cells by TLR ligation

doi: 10.1186/1475-2867-14-54

Figure Lengend Snippet: Sequence, amplicon size and annealing temperatures of primers used in this study

Article Snippet: The membranes were blocked with 5% non-fat dry milk and probed with 1:100 dilution of goat anti-human TLR2 (Santa Cruz, USA) or 1 μg/mL goat anti-human TLR4 (R&D, USA) for 2 h. Membranes were consequently incubated with peroxidase-conjugated rabbit anti-goat secondary antibody (Avicenna research Institute, Iran) at a dilution of 1:10000.

Techniques: Sequencing, Amplification