Journal: Nature Communications

Article Title: Lactobacillus crispatus S-layer proteins modulate innate immune response and inflammation in the lower female reproductive tract

doi: 10.1038/s41467-024-55233-7

Figure Lengend Snippet: HEK TLR2 ( a ) and VK2 vaginal epithelial cells ( b ) were incubated with bacteria (MOI = 10) overnight. TLR1 and TLR6-dependence was investigated by pre-incubating the cells for 30 min with 1 μg/ml anti-TLR1, anti-TLR6 or mIgG1 isotype control antibodies before overnight stimulation with the bacteria at MOI = 10. Fv Fannyhessea vaginae , Fn Fusobacterium nucleatum , Pb Prevotella bivia , Sv Sneathia vaginalis , Mm Mobiluncum mulieris , Fm Fingoldia magna , Va Veillonella atypica . Data are mean +/−SD ( n = 3 biological replicates). Two-way ANOVA, Tuckey multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0001. Source data are provided as a Source Data file.

Article Snippet: To investigate TLR1, TLR2 and TLR6 dependence, VK2/E6E7 cells were pre-incubated for 30 min at 37 °C with 1 μg/ml of anti-hTLR1 (mabg-htlr1, clone H2G2, InvivoGen), anti-hTLR2 (MAB2616, clone # 383936, R&D Systems) and anti-hTLR6 (mabg-htlr6, clone C5C8, InvivoGen) antibody or mIgG1 (InvivoGen, clone T8E5, mabg1-ctrlm) and mIgG2b (14-4732-85, lot 2288614, clone eBMG2b, ThermoFisher Scientific) isotype controls.

Techniques: Incubation, Bacteria, Control, Comparison

Journal: Nature Communications

Article Title: Lactobacillus crispatus S-layer proteins modulate innate immune response and inflammation in the lower female reproductive tract

doi: 10.1038/s41467-024-55233-7

Figure Lengend Snippet: a SDS-PAGE gel of crude SLPs extracted with LiCl 5 M from L. crispatus and L. iners isolates. 10μg total proteins were loaded per lane. Representative of three independent experiments. b HEK-TLR2 cells were stimulated for 16 h with bacteria (MOI = 10) and NF-κB activation was determined by measuring alkaline phosphatase activity and reading O.D. at 630 nm. c VK2 vaginal epithelial cells were stimulated for 16 h and IL-8 release in the supernatant was quantified by ELISA. d HEK TLR2 cells were stimulated with lithium chloride-treated bacteria (MOI = 10) for 16 h and IL-8 release in the supernatant was quantified by ELISA. TLR1, TLR2 and TLR6 dependence was investigated by pre-incubating cells for 30 min at 37 °C with 1 μg/ml of anti-TLR1, anti-TLR2, anti TLR6 or mIgG1 and mIgG2b isotype control antibodies. b – d A representative figure of three independent experiments is shown (mean +/− SD). Two-way ANOVA, Tuckey multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.005, **** p < 0,001. Source data are provided as a Source Data file.

Article Snippet: To investigate TLR1, TLR2 and TLR6 dependence, VK2/E6E7 cells were pre-incubated for 30 min at 37 °C with 1 μg/ml of anti-hTLR1 (mabg-htlr1, clone H2G2, InvivoGen), anti-hTLR2 (MAB2616, clone # 383936, R&D Systems) and anti-hTLR6 (mabg-htlr6, clone C5C8, InvivoGen) antibody or mIgG1 (InvivoGen, clone T8E5, mabg1-ctrlm) and mIgG2b (14-4732-85, lot 2288614, clone eBMG2b, ThermoFisher Scientific) isotype controls.

Techniques: SDS Page, Bacteria, Activation Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Control, Comparison