Journal: Journal of Virology

Article Title: GB Virus B and Hepatitis C Virus, Distantly Related Hepaciviruses, Share an Entry Factor, Claudin-1

doi: 10.1128/jvi.00469-23

Figure Lengend Snippet: Extracellular loop 2 of claudin-1 is important for GBV-B entry. (A and B) Huh7 CLDN1 KO (A) or HEK293T (B) cells transduced to express CLDN1, CLDN9, CLDN6, or GFP were challenged with pseudotyped viruses harboring glycoproteins from GBV-B, HCV J6, HCV H77, and VSV. (C) Claudin-1 (black) and claudin-9 (gray) topologies with the indicated regions of each protein swapped with the respective regions of the other CLDNs to produce chimeric proteins, resulting in chimeric proteins with the following amino acids: (CLDN1 aa 1 to 139/CLDN9 aa 138 to 217) (ChimA), CLDN9 aa 1 to 81/CLDN1 aa 82 to 211 (ChimB), CLDN9 aa 1 to 137/CLDN1 aa 139 to 211 (ChimC), and CLDN1 aa 1 to 81/CLDN9 aa 82 to 217 (ChimD). (D and F) Chimeric protein expression in HEK293T cells (D) or Huh7 cells (F) was assessed by immunoblotting using anti-CLDN1 and anti-CLDN9 antibodies. The protein input was verified using an anti-actin antibody. (E) HEK293T cells transduced to express the indicated chimeric CLDN1/9 proteins were infected with H77pp. (G) Huh7 CLDN1 KO cells transduced to express the indicated chimeric CLDN1/9 proteins were infected with GBVBpp, J6pp, J77pp, and VSVpp. Infectivity is expressed as the mean RLU values ± standard deviations from 2 independent experiments run in triplicate. *, P < 0.05; **, P < 0.005; ***, P < 0.0005 (by a Kruskal-Wallis test compared to the wild type [WT]).

Article Snippet: Membranes were probed by serial incubation with rabbit anti-CLDN1 antibody (1:1,000; Abcam) or rabbit anti-CLDN9 antibody (1:300; Proteintech), rabbit anti-β-actin antibody (1:10,000; Abcam), and goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (HRP).

Techniques: Expressing, Western Blot, Infection

Journal: Journal of Virology

Article Title: GB Virus B and Hepatitis C Virus, Distantly Related Hepaciviruses, Share an Entry Factor, Claudin-1

doi: 10.1128/jvi.00469-23

Figure Lengend Snippet: Overlap extension PCR primers

Article Snippet: Membranes were probed by serial incubation with rabbit anti-CLDN1 antibody (1:1,000; Abcam) or rabbit anti-CLDN9 antibody (1:300; Proteintech), rabbit anti-β-actin antibody (1:10,000; Abcam), and goat anti-rabbit secondary antibody conjugated to horseradish peroxidase (HRP).

Techniques: Sequencing

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA alleviates neuropathic pain and enhances intestinal barrier function. (A) The concentration of total bile acids in fecal samples (n = 6). (B) The expression of HDCA in fecal samples (n = 6). (C) Schedule for the management of HDCA. (D, E) PWT and PWL of mice following HDCA treatment (n = 6). Representative immunoblots (F) and relative expression levels of FXR mRNA (G) and FXR protein (H). (I) Immunohistochemical staining of FXR + cells and Alcian Blue staining in the distal ileum (scale bar, 100 μm, n = 6). (J) Quantitative analysis of FXR + cells. The relative expression levels of ZO-1 (K), occludin (L) and claudin-1 (M) protein in distal ileum (n = 3). (N) Representative immunoblots and relative expression levels of Mucin-2 (P) (n = 3). (O) Quantitative analysis levels of the number of goblet cells in the distal ileum (n = 6). (Q, R) Immunofluorescence staining and mean fluorescence intensity of Mucin-2 (scale, 100 μm, n = 3). (S) Transmission electron microscopy examination of the distal ileum. (scale, 1.0 μm). * P < 0.05, ** P < 0.01 by unpaired Student's t‐test (A and B). * P < 0.05, ** P < 0.01 *** P < 0.001, # P < 0.05, ## P < 0.01 by Two-way repeated ANOVA with post hoc Bonferroni’s test (D and E), * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (G, H, J–M, O–Q). Data is presented as mean ± SEM. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Membranes were blocked using 5 % skim milk at room temperature for 1 h and subsequently incubated at 4 °C overnight with primary antibodies against FXR (1:1000, Immunoway, YN2161), zonula occludens-1 (ZO-1) (1:1000, Affinity, AF5145), Mucin-2 (1:1000, Abcam, ab272692), occludin (1:1000, Proteintech, 27260–1-AP), claudin-1 (1:1000, WL, WL03073), MMP-2 (1:1000, Immunoway, YT2798), MMP-9 (1:1000, Immunoway, YT1892), PPAR-γ (1:1000, Proteintech, 16643-1-AP), NOD-like receptor protein-3 (NLRP3) (1:1000, Immunoway, YT5382), interleukin-6 (IL-6) (1:1000, WL, WL02841), IL-18 (1:1000, WL, WL01127), tumor necrosis factor-alpha (TNF-α) (1:1000, WL, WL01581), IL-10 (1:1000, WL, WL03088), FGF15 (1:500, Santacruz, sc-514647), TGR5 (1:1000, Abcam, ab72608), BSEP (1:1000, Abcam, ab155421), CYP7α1 (1:1000, Abcam, ab65596) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:5000, Proteintech, 60004-1-Ig).

Techniques: Concentration Assay, Expressing, Western Blot, Immunohistochemical staining, Staining, Immunofluorescence, Fluorescence, Transmission Assay, Electron Microscopy