Journal: mBio

Article Title: Chlamydia trachomatis modulates the expression of JAK-STAT signaling components to attenuate the type II interferon response of epithelial cells

doi: 10.1128/mbio.01834-24

Figure Lengend Snippet: IFNγ-induced activation of STAT1 is attenuated in C. trachomatis L2-infected Hep2 cells. ( A ) Schematic representation of C. trachomatis serovar L2 infection and IFNγ treatment of epithelial HEp2 cells. ( B ) C. trachomatis -infected cells (multiplicity of infection [MOI] = 2, yellow outline for cell boundary as determined in bright-field channel), chlamydial inclusion visualized with human anti- C . trachomatis antibody in cyan) and uninfected HEp2 cells exposed to IFNγ (10 U/mL) starting at 24 hpi for 6 hours were fixed and stained for phosphorylated STAT1 (pSTAT1) in red and for DNA with DAPI in blue. ( C ) Nuclear pSTAT1 fluorescence intensities were quantified in both uninfected and infected cells. Measurements were normalized to respective no IFNγ control to account for background pSTAT1 staining and plotted as mean fluorescence. Violin plot represents six independent experiments with total of 260 nuclei measured per group. Statistical significance was determined by Wilcoxon rank sum. ( D ) Protein level of IFNγ-STAT1 downstream target IFO1 was assessed from total protein lysates collected from C. trachomatis -infected (MOI = 4) and uninfected cells exposed to IFNγ (10 U/mL) at 24 hpi for 6 hours. ( E ) Densitometric quantification of IDO1 plotted as ratios to the loading control α-tubulin. Bar graph represents four independent experiments, and statistical significance was determined using Welch’s t -test. * P < 0.05 and *** P < 0.001. Scale bar at 10 µm.

Article Snippet: HEp2 cells were seeded onto 12-mm acid-etched glass coverslips (Chemglass, CLS-1760-012) placed in 24-well plates.

Techniques: Activation Assay, Infection, Staining, Fluorescence, Control

Journal: mBio

Article Title: Chlamydia trachomatis modulates the expression of JAK-STAT signaling components to attenuate the type II interferon response of epithelial cells

doi: 10.1128/mbio.01834-24

Figure Lengend Snippet: C. trachomatis growth is inhibited in cells pretreated with IFNγ. Immunofluorescence visualization of C. trachomatis inclusions using the chlamydial protein cHsp60 in HEp2 cells following pre-treatment (Pre-12 + 24) with IFNγ for 12 hours before infection or co-treatment (Co-24) with IFNγ at time of infection. Scale bar at 20 µm.

Article Snippet: HEp2 cells were seeded onto 12-mm acid-etched glass coverslips (Chemglass, CLS-1760-012) placed in 24-well plates.

Techniques: Immunofluorescence, Infection

Journal: PLOS Pathogens

Article Title: Isolation and characterization of IgG3 glycan-targeting antibodies with exceptional cross-reactivity for diverse viral families

doi: 10.1371/journal.ppat.1012499

Figure Lengend Snippet: (A) HEp-2 cell slides were immunofluorescently stained with the indicated mAbs and goat anti-human Ig-FITC secondary (green) and DAPI (blue, to stain cell nuclei). Each mAb was tested at 100, 10, and 1 μg/mL. Control human serum samples that were positive or negative for anti-nuclear antibodies are shown (right). Images are 40X magnification. (B) Autoantigens were tested for reactivity by 2526, Ab82 (negative control antibody), and A32 (positive control antibody) using a commercially available AtheNA Multi-Lyte ANA kit. Values shown in bold were considered positive.

Article Snippet: HEp-2 cell coated slides (BION ENTERPRISES LTD ANA (HEp-2) Test System, ANK-120) were incubated with purified antibodies at 100, 10, or 1 μg/ml or control sera in a moist chamber at room temperature for 30 min. Controls provided with the kit included anti-nuclear antibody (ANA)+ and ANA- human sera.

Techniques: Staining, Control, Negative Control, Positive Control