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MRL/lpr mice were intraperitoneally injected weekly with ( A ) PBS or ( B ) curli purified from Salmonella Typhimurium for 6 weeks and euthanized at 13 weeks old. Frozen tissue sections were stained with <t>goat</t> <t>anti-mouse</t> IgG-AlexaFluor405 (blue), rat anti-mouse C3-AlexaFluor647 (red), and rat anti-mouse C1q-FITC (green). ( A-B ) Representative images of glomeruli by confocal microscopy (63X objective). Sections shown are from two individual mice per group, scale bars represent 20 µm. ( C-F ) Images were quantified in ImageJ for mean fluorescence intensity (MFI) inside glomeruli of ( C ) IgG, ( D ) C3, ( E ) C1q, and ( F ) area was measured, within at least 15 glomeruli per mouse and averaged. Statistics: ( C-F ) Unpaired t-test (**p < 0.01).
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MRL/lpr mice were intraperitoneally injected weekly with ( A ) PBS or ( B ) curli purified from Salmonella Typhimurium for 6 weeks and euthanized at 13 weeks old. Frozen tissue sections were stained with <t>goat</t> <t>anti-mouse</t> IgG-AlexaFluor405 (blue), rat anti-mouse C3-AlexaFluor647 (red), and rat anti-mouse C1q-FITC (green). ( A-B ) Representative images of glomeruli by confocal microscopy (63X objective). Sections shown are from two individual mice per group, scale bars represent 20 µm. ( C-F ) Images were quantified in ImageJ for mean fluorescence intensity (MFI) inside glomeruli of ( C ) IgG, ( D ) C3, ( E ) C1q, and ( F ) area was measured, within at least 15 glomeruli per mouse and averaged. Statistics: ( C-F ) Unpaired t-test (**p < 0.01).
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MRL/lpr mice were intraperitoneally injected weekly with ( A ) PBS or ( B ) curli purified from Salmonella Typhimurium for 6 weeks and euthanized at 13 weeks old. Frozen tissue sections were stained with <t>goat</t> <t>anti-mouse</t> IgG-AlexaFluor405 (blue), rat anti-mouse C3-AlexaFluor647 (red), and rat anti-mouse C1q-FITC (green). ( A-B ) Representative images of glomeruli by confocal microscopy (63X objective). Sections shown are from two individual mice per group, scale bars represent 20 µm. ( C-F ) Images were quantified in ImageJ for mean fluorescence intensity (MFI) inside glomeruli of ( C ) IgG, ( D ) C3, ( E ) C1q, and ( F ) area was measured, within at least 15 glomeruli per mouse and averaged. Statistics: ( C-F ) Unpaired t-test (**p < 0.01).
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MRL/lpr mice were intraperitoneally injected weekly with ( A ) PBS or ( B ) curli purified from Salmonella Typhimurium for 6 weeks and euthanized at 13 weeks old. Frozen tissue sections were stained with goat anti-mouse IgG-AlexaFluor405 (blue), rat anti-mouse C3-AlexaFluor647 (red), and rat anti-mouse C1q-FITC (green). ( A-B ) Representative images of glomeruli by confocal microscopy (63X objective). Sections shown are from two individual mice per group, scale bars represent 20 µm. ( C-F ) Images were quantified in ImageJ for mean fluorescence intensity (MFI) inside glomeruli of ( C ) IgG, ( D ) C3, ( E ) C1q, and ( F ) area was measured, within at least 15 glomeruli per mouse and averaged. Statistics: ( C-F ) Unpaired t-test (**p < 0.01).

Journal: PLOS Pathogens

Article Title: Salmonella infection and biofilms drive anti-C1q and anti-dsDNA autoantibodies in systemic autoimmunity

doi: 10.1371/journal.ppat.1014666

Figure Lengend Snippet: MRL/lpr mice were intraperitoneally injected weekly with ( A ) PBS or ( B ) curli purified from Salmonella Typhimurium for 6 weeks and euthanized at 13 weeks old. Frozen tissue sections were stained with goat anti-mouse IgG-AlexaFluor405 (blue), rat anti-mouse C3-AlexaFluor647 (red), and rat anti-mouse C1q-FITC (green). ( A-B ) Representative images of glomeruli by confocal microscopy (63X objective). Sections shown are from two individual mice per group, scale bars represent 20 µm. ( C-F ) Images were quantified in ImageJ for mean fluorescence intensity (MFI) inside glomeruli of ( C ) IgG, ( D ) C3, ( E ) C1q, and ( F ) area was measured, within at least 15 glomeruli per mouse and averaged. Statistics: ( C-F ) Unpaired t-test (**p < 0.01).

Article Snippet: Tissue was incubated in antibody cocktails including 1:30 goat anti-mouse IgG conjugated to AlexaFluor 405 (Invitrogen A31553), 1:25 rat anti-mouse C3 (11H9) conjugated to AlexaFluor 647 (Santa Cruz Biotechnology sc-58926), and 1:20 rat anti-mouse C1q (7H8) conjugated to FITC (Santa Cruz Biotechnology sc-58920) diluted in PBS-T with 1% bovine serum albumin for 2 hours at room temperature, covered with a small circle of parafilm and kept in the dark underneath a lid to prevent evaporation and bleaching.

Techniques: Injection, Purification, Tissue, Staining, Confocal, Microscopy, Fluorescence