Journal: One Health

Article Title: Human 3D liver spheroids support productive infection of a novel tick-borne phenuivirus

doi: 10.1016/j.onehlt.2026.101321

Figure Lengend Snippet: Adaptation and pathogenesis of MKWV in human 3D liver spheroids. (A) Schematic of serial passaging of the HLJ1 strain in spheroids, yielding the adapted NAC-Org5 strain. (B, C) Viral RNA copies (B) and TCID₅₀ titers (C) across passages (P1-P5). (D) Bright-field image of spheroids infected with passage 5 (P5) virus, showing structural disruption. Scale bar, 100 μm. (E) Quantification of spheroid diameter post-infection. (F) Transmission electron micrographs of virions within cytoplasmic vesicles of infected spheroids. Scale bars: 1 μm (left), 200 nm (right). (G) Representative images and quantification of nuclei showing infection-induced cell death. Scale bar, 200 μm. (H) Western blot detecting cleaved caspase-3 in spheroids at 48 and 72 h post-infection (hpi). (I) Multiplex immunofluorescence showing NAC-Org5 tropism for CD31 + endothelial cells and CD68 + Kupffer cells, with weaker detection in ALB + hepatocytes. Scale bar, 200 μm. (J) Functional assessment of infected spheroids: ATP (viability), ALT/AST/LDH (damage), ALB/urea (synthetic function). (K) RT-qPCR analysis of pro-inflammatory cytokine mRNA expression, normalized to β-actin. Data are mean ± SD ( n = 5 biological replicates). * p < 0.05, ** p < 0.01.

Article Snippet: After blocking with 5% non-fat milk, membranes were incubated with anti- Caspase-3 antibody (Cell Signaling Technology, 144220, 1:1000) followed by HRP-conjugated secondary antibody.

Techniques: Passaging, Infection, Virus, Disruption, Transmission Assay, Western Blot, Multiplex Assay, Immunofluorescence, Functional Assay, Quantitative RT-PCR, Expressing

Journal: Cancer letters

Article Title: WEE1 Stabilizes MYC to Promote Therapeutic Resistance in Esophageal Adenocarcinoma

doi: 10.1016/j.canlet.2026.218418

Figure Lengend Snippet: A. 21 positive hits (red), which synergized with MK-1775 in O19 cells during the first round of screening using an 892 FDA-approved drug screening library. B. Panobinostat (red arrow) was found to synergize with MK-1775 in FLO1 (blue) and SK-GT4 (red) cell lines during the second round of screening. C. Synergy Finder analysis results in FLO1, OE33, OE19, and SK-GT4 cell lines treated with MK-1775 and Panobinostat. *P<0.05, **P<0.01, ***P<0.001. D. Flow cytometric analysis of Annexin V & SYTOX red dual staining in OE33 cells treated with MK-1775/Panobinostat alone or a combination. E. Percentage of apoptotic cells from Fig D *P<0.05, **P<0.01, ***P<0.001. F. Western blot analysis of PARP, Cl-PARP, Caspase 3, Cl-Caspase 3, and β-ACTIN in OE33 cells treated with MK-1775/ Panobinostat alone or a combination. G. Flow cytometric analysis of Annexin V & SYTOX red dual staining in OE19 cells treated with MK-1775/Panobinostat alone or a combination. H. Percentage of apoptotic cells from Fig G *P<0.05, **P<0.01, ***P<0.001. I. Western blot analysis of PARP, Cl-PARP, Caspase 3, Cl-Caspase 3, and β-ACTIN in OE19 cells treated with MK-1775/ Panobinostat alone or a combination.

Article Snippet: C-MYC, p-C-MYC T58, P-GSK3β S9, GSK3β, PARP, CL-PARP, Caspase 3, Cl-Caspase 3, HA tag, CD133, SOX9, SOX2, CD44, OCT4, Nanog and Ki-67 antibodies were obtained from Cell Signaling Technologies (Danvers, MA).

Techniques: Drug discovery, Staining, Western Blot

Journal: Cancer letters

Article Title: WEE1 Stabilizes MYC to Promote Therapeutic Resistance in Esophageal Adenocarcinoma

doi: 10.1016/j.canlet.2026.218418

Figure Lengend Snippet: A. Human EAC PDX-derived Organoids treated with MK-1775 / Panobinostat alone or in combination. B. Quantification of organoid diameter from A. C. Tumor growth curves in 4 experimental groups (Untreated control, MK-1775, Panobinostat, combination of MK-1775 & Panobinostat) at the end of the experiment. D. Western blot analysis of PARP, Cl-PARP, Caspase 3, CL-Caspase3, and β-ACTIN in Mouse Xenografts. E. Immunofluorescence staining for P-CDC2 Y15 (green), C-MYC (red), MRP1 (green), and Ki67 (red) in Mouse EAC PDXs, captured at 20X Magnification. F. Quantification data from E. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: C-MYC, p-C-MYC T58, P-GSK3β S9, GSK3β, PARP, CL-PARP, Caspase 3, Cl-Caspase 3, HA tag, CD133, SOX9, SOX2, CD44, OCT4, Nanog and Ki-67 antibodies were obtained from Cell Signaling Technologies (Danvers, MA).

Techniques: In Vivo, Derivative Assay, Control, Western Blot, Immunofluorescence, Staining

Journal: Journal of Human Immunity

Article Title: Human ISG15 deficiency unveils impaired healing of ulcerations via type I interferon–mediated fibrosis

doi: 10.70962/jhi.20250011

Figure Lengend Snippet: IFNα induces apoptosis in ISG15 KO cells. (A) Healing/migration ability of WT and ISG15 KO fibroblasts was assessed with the scratch assay. Left; percent confluency was recorded every hour for a total of 93 h. Right; experimental replicates at 48, 72, and 93 h after scratch. (B) Representative images of WT and ISG15 KO fibroblasts at 1 h (left), 49 h (middle), and 92 h (right) of 1,000 IU ml −1 IFNα treatment. White arrows indicate macroscopic evidence of cellular stress. (C) WT and ISG15 KO fibroblasts were treated with IFNα2b (0, 100, 1,000 IU/ml) for 72 h, and cell death was quantified by measuring Annexin V and Propidium Iodide (PI) levels through flow cytometry. (D) Control and ISG15 KO fibroblasts were treated with IFNα2b (0, 100, 1,000 IU/ml) for 72 h following a 30-min pretreatment with necrostatin-1, and cell death was quantified by measuring Annexin V and PI levels through flow cytometry. (E) Metabolic activity of two control and one ISG15 KO fibroblast cell lines was assessed with the Deep Blue assay. Nec-1; necrostatin-1. (F) Control and ISG15-deficient fibroblasts were treated with either IFNα2b (1, 100, 1,000 IU/ml) or chloroquine (0, 25, 50, 100 μM) for 72 h. Cell lysates were analyzed by western blotting for autophagy (LC3B). (G) Control and ISG15-deficient fibroblasts were treated with either IFNα2b (100 IU/ml), or TNFα (5 ng/μl), or both for 72 h, and percent death was assessed with the Deep Blue assay as compared to the nontreated condition for each variant. (H) WT and ISG15 KO fibroblasts were treated with IFNα2b (0, 100, 1,000 IU/ml) for 72 h, and cell death was quantified by measuring cleaved (active) caspase 3 levels through flow cytometry. Source data are available for this figure: . P values were calculated with two-tailed t test. **P < 0.01; ***P < 0.001.

Article Snippet: The presence of caspase 3 was probed with anti-caspase 3 antibody (Cat. PP229AA; Biocare Medical).

Techniques: Migration, Wound Healing Assay, Flow Cytometry, Control, Activity Assay, Western Blot, Variant Assay, Two Tailed Test

Journal: Journal of Human Immunity

Article Title: Human ISG15 deficiency unveils impaired healing of ulcerations via type I interferon–mediated fibrosis

doi: 10.70962/jhi.20250011

Figure Lengend Snippet: Reconstitution with WT ISG15 and inhibition of apoptosis rescue apoptosis of IFNα-treated ISG15 KO cells. (A) Healing/migration ability of WT, UBE1L KO, and ISG15 KO fibroblasts, as well as ISG15 KO cells complemented with WT ISG15 or Luc, was assessed with the scratch assay for 93 h. (B) Apoptosis following 72-h IFNα2b treatment of WT, UBE1L KO, and ISG15 KO fibroblasts, as well as ISG15 KO cells complemented with WT ISG15 or Luc, was assessed by quantifying cleaved caspase 3 levels through flow cytometry. (C) Rescue of A549 ISG15 KO IFNα2b-induced cell death with pan-caspase inhibitor (Z-VAD). (D) Proliferation of ISG15 KO lung epithelial cells (A549) with or without IFNα2b as compared to control. (E) CASP3 IHC for three healthy volunteer and three patient’s skin biopsies. White arrows indicate areas of cleaved Caspase-3 (brown).

Article Snippet: The presence of caspase 3 was probed with anti-caspase 3 antibody (Cat. PP229AA; Biocare Medical).

Techniques: Inhibition, Migration, Wound Healing Assay, Flow Cytometry, Control

Journal: International Journal of Molecular Medicine

Article Title: IL-37/IL-1R8 blocks keratinocyte acantholysis via suppressing ADAM17/EGFR

doi: 10.3892/ijmm.2026.5793

Figure Lengend Snippet: IL-37 protects HaCaT cells from keratinocyte dissociation and apoptosis. (A) The effect of IL-37 on cell dissociation was evaluated using a cell dissociation assay. (B) Mitochondrial membrane potential was assessed by JC-1 staining. (C) Cell apoptosis was analyzed by flow cytometry. (D) Protein expression levels of Bcl-2, Bax, Cleaved-caspase 3 and Caspase 3 were determined by western blotting. Quantitative analysis of (E) Bcl-2, (F) Bax and (G) Cleaved-caspase 3 protein levels. * P<0.05 vs. Control group; # P<0.05 vs. anti-Dsg3 group. Data are presented as mean ± SD, n=3 biological independent replicates. One-way ANOVA with Bonferroni's post-hoc test was used for multiple group comparisons. IL, interleukin.

Article Snippet: Following this, the membranes were incubated with the following specific primary antibodies overnight at 4°C: Rabbit anti-IL-37 monoclonal antibody (cat. no. ab278499; Abcam), rabbit anti-Bax monoclonal antibody (cat. no. ab32503; Abcam), rabbit anti-Bcl-2 polyclonal antibody (cat. no. ab59348; Abcam), rabbit anti-caspase 3 antibody (cat. no. 9662; Cell Signaling Technology, Inc.), rabbit anti-cleaved caspase3 antibody (cat. no. 9661; Cell Signaling Technology, Inc.), mouse anti-IL-1R8 antibody (cat. no. sc-271864; Santa Cruz Biotechnology, Inc.), mouse anti-IL-18Rα antibody (cat. no. PA5-115404; Thermo Fisher Scientific, Inc.), rabbit anti-EGFR phospho Y1173 antibody (cat. no. ab5652; Abcam), rabbit anti-EGFR antibody (cat. no. ab52894; Abcam), rabbit anti-ADAM17 polyclonal antibody (cat. no. ab39162; Abcam), anti-phosphorylated (p-) ERK1/2 antibody (cat. no. ab201015; Abcam), anti-ERK1/2 antibody (cat. no. ab184699; Abcam), anti-p-AKT (Ser473) antibody (cat. no. 9271; Cell Signaling Technology, Inc.), anti-AKT antibody (cat. no. 4691; Cell Signaling Technology, Inc.), anti-p-STAT3 antibody (cat. no. ab76315; Abcam), and anti-STAT3 antibody (cat. no. ab68153; Abcam).

Techniques: Membrane, Staining, Flow Cytometry, Expressing, Western Blot, Control