Journal: Cell reports

Article Title: MyD88 signaling in myeloid cells induces gastrointestinal tract injury and systemic inflammation after West Nile virus infection

doi: 10.1016/j.celrep.2025.116852

Figure Lengend Snippet: WT C57BL/6J mice were treated with anti-IFNAR1 mAb plus isotype control or anti-CCR2 mAb (25 μg/mouse) and subcutaneously inoculated with 10 2 FFU of WNV 1 day later. Two additional doses of isotype control or anti-CCR2 mAb were administered at 1 and 3 dpi. At 5 dpi, mice were administered fluorescently conjugated dextrans by oral gavage. (A-B and E-F) Confocal microscopy images of duodenal sections at 5 dpi. (A) WNV antigen (red), monocytes (CCR2, green), EpCAM (white), and nuclei (Hoechst 33258, blue). Scale bars, 100 μm (upper panel) or 50 μm (lower panel). (B) Quantification of CCR2-positive cells in the GI tract, expressed as percentage of total Hoechst 33258-positive cells per field. (C and D) Serum levels of 10 (C) and 250 (D) kDa dextran. (E and F) Quantification of WNV antigen-positive cells in the GI tract, expressed as percentage of total Hoechst 33258-positive cells per field (E) or as percentage of total EpCAM-positive epithelial cells per field (F). (G and H) Hematoxylin and eosin staining of liver sections from mice at 5 dpi. Scale bars, 100 μm (G). (H) Quantitation of liver injury. (I–K) Confocal microscopy images of cerebral cortex sections at 5 dpi showing Iba1 (white) and nuclei (Hoechst 33258, blue). Scale bars, 100 μm (upper panel) or 50 μm (lower panel) (I). (J) Iba1-positive area was quantified from at least 15 microglia/macrophage cells per field and normalized to the number of microglia/macrophage nuclei counted. (K) Numbers of Iba1-positive microglia/macrophages per field. Each data point is derived from at least three independent fields per mouse brain (J–K). (L–O) Cytokine levels of TNF (L and M) and IL-6 (N and O) in serum (L and N) and brain homogenates (M and O). (P–S) WNV RNA levels in the serum (P), spleen (Q), liver (R), and brain (S) at 5 dpi. Bars indicate median values from two experiments; dotted lines show LODs; n = 10 mice (A–F and I–S); n = 6 mice (G–H). Statistical analysis: Mann-Whitney test (ns: non-significant, ** p < 0.01, *** p < 0.001, and **** p < 0.0001).

Article Snippet: Sections were incubated overnight at 4°C with rat anti-WNV hyperimmune serum (1:750 dilution), rabbit anti-EpCAM polyclonal serum (1:2000, Abcam) and goat anti-Iba1 polyclonal serum (1:500, ThermoFisher) or rabbit anti-CCR2 polyclonal serum (1:500, Novus Biologicals) diluted in 1X TBS containing 1% BSA, 3% normal donkey serum, and 0.1% Triton X-100.

Techniques: Control, Confocal Microscopy, Staining, Quantitation Assay, Derivative Assay, MANN-WHITNEY

Journal: Iranian Journal of Microbiology

Article Title: Yeast-mediated display: probing Helicobacter pylori HopQ and CEACAM1 interaction

doi: 10.18502/ijm.v17i6.20366

Figure Lengend Snippet: SDS-PAGE and Western blot analyses. In panel A, the left image shows Coomassie Blue staining of S. cerevisiae expressing Aga2-C1ND (lanes 1 and 2). The middle blot shows two distinct bands at 50 kDa and 24 kDa detected using an HRP-conjugated anti-GFP antibody. Lane 1 corresponds to S. cerevisiae expressing the GFP-C1ND fusion construct. The presence of a 27 kDa band suggests cleavage of EGFP from its fusion partner. The right image shows signals detected by the anti-C1ND antibody for non-EGFP-tagged C1ND. The 24 kDa band corresponds to the Aga2-C1ND fusion protein. In panel B, the left image shows positive signals detected using the anti-C1ND antibody. Lane 1 shows the expression of the GFP-C1ND fusion protein, with bands at 24 kDa and 50 kDa. Lanes 2 and 3 represent strains expressing the Aga2-C1ND fusion protein, showing a band at 24 kDa. The positive control (lane 4) expresses the intact C1ND fragment, displaying a band at 12 kDa. The right image shows detection of the EGFP-C1ND fusion protein (50 kDa, lane 1) in a positive transformant using an HRP-conjugated anti-His-tag antibody. The positive control in this blot (lane 2) corresponds to an irrelevant His-tagged protein. In all blots, “C−“ indicates non-transformed lysate. Blots in panels A and B were developed using ECL and DAB staining, respectively.

Article Snippet: Immunoblots were probed overnight at 4°C with HRP-labeled antibodies against GFP (1:2000; Acris, USA), His-tag (1:1000; Sigma, USA), and C1ND (1:3000; Sigma, USA).

Techniques: SDS Page, Western Blot, Staining, Expressing, Construct, Positive Control, Transformation Assay

Journal: Iranian Journal of Microbiology

Article Title: Yeast-mediated display: probing Helicobacter pylori HopQ and CEACAM1 interaction

doi: 10.18502/ijm.v17i6.20366

Figure Lengend Snippet: Confocal microscopy images showing the surface expression of the Aga2-EGFP-G4S-C1ND fusion protein in S. cerevisiae , indicated by arrows. (A) Green fluorescence from the EGFP tag. (B) Red surface fluorescence of recombinant yeast cells stained with PE-conjugated anti-GFP antibody. (C) Merged image of panels A and B. (D) Brightfield image. (E) No fluorescence in the negative control. Scale bar: 5 μm

Article Snippet: Immunoblots were probed overnight at 4°C with HRP-labeled antibodies against GFP (1:2000; Acris, USA), His-tag (1:1000; Sigma, USA), and C1ND (1:3000; Sigma, USA).

Techniques: Confocal Microscopy, Expressing, Fluorescence, Recombinant, Staining, Negative Control