Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: The aryl hydrocarbon receptor (AHR) drives human leukocyte antigen (HLA)-II expression in human melanoma

doi: 10.1186/s13046-026-03673-y

Figure Lengend Snippet: AHR and ARNT regulate basal HLA-II expression on cancer cells. A Western blot analysis confirming AHR or ARNT overexpression (OE) in A375, WM115, and SKMEL2 melanoma cell lines. Protein levels of AHR, ARNT, and HLA-DRA were assessed using anti-AHR (clone D5S6H), anti-ARNT (clone D28F3), and polyclonal anti–HLA-DRA antibodies, respectively, with β-tubulin detected using an anti–β-tubulin antibody (clone C66) as a loading control. B- C Flow cytometry analysis of surface pan–HLA-II expression in A375, WM115, and SKMEL2 cells overexpressing AHR or ARNT. Representative histograms from three independent experiments show pan–HLA-II expression, with dashed lines indicating the median of the empty vector control group ( B ). The gMFI was quantified across three independent experiments, normalized to the empty vector control group, and presented as fold change ( C ). D Western blot analysis of AHR, ARNT, and HLA-DRA protein levels in AHR or ARNT reconstituted A375 cells generated by reintroducing AHR or ARNT into respective KO cells. Protein levels of AHR, ARNT, and HLA-DRA were assessed using anti-AHR (clone D5S6H), anti-ARNT (clone D28F3), and polyclonal anti–HLA-DRA antibodies, respectively, with β-tubulin detected using an anti–β-tubulin antibody (clone C66) as a loading control. E - F Flow cytometry analysis of surface pan–HLA-II expression in AHR- or ARNT-reconstituted A375 cells. Representative histograms from three independent experiments show pan–HLA-II expression ( E ). The gMFI was quantified across three independent experiments, normalized to the NTC group, and presented as fold change ( F ). Surface pan–HLA-II expression was detected using APC anti-human HLA-DR, DP, DQ Antibody (clone Tü39) ( B - C and E - F ). Data are represented as mean ± SD ( C and F ). Statistical analysis by one-way ANOVA ( C ) and unpaired Student’s t-test ( F ); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. vec, empty vector control

Article Snippet: The following primary antibodies were used: AHR Rabbit mAb (clone D5S6H, CST #83200, RRID: AB_2800011), ARNT Rabbit mAb (clone D28F3, CST #5537, RRID: AB_10694232), HLA-DRA polyclonal Rabbit Ab (Boster #A01195), β-Tubulin Mouse mAb (clone C66, Abmart # M20005 , RRID: AB_2920648), and β-Actin Rabbit mAb (clone 13E5, CST #4970, RRID: AB_2223172).

Techniques: Expressing, Western Blot, Over Expression, Control, Flow Cytometry, Plasmid Preparation, Generated

Journal: Bioactive Materials

Article Title: Hypoxia preconditioned MSC exosomes attenuate high-altitude cerebral edema via the miR-125a-5p/RTEF-1 axis to protect vascular endothelial cells

doi: 10.1016/j.bioactmat.2025.06.018

Figure Lengend Snippet: Identification and characterization of H-EXO. (A) Extraction schematic of N-EXO and H-EXO (created with biorender.com ). (B) Morphological characteristics of N-EXO and H-EXO under transmission electron microscope. (C), (D) NTA technique was used to detect N-EXO and H-EXO. (E) Dil labels phagocytic exosomes in vascular endothelial cells. (F) To investigate the expression of exosome-specific protein markers CD9, CD63, CD81, Calnexin and GM130 by WB. (G) Protein concentrations of N-EXO and H-EXO. All data are presented as mean ± SD; Scale bars: 1 μm (B left), 500 nm (B right), 573 nm (D), 25 μm (E left), 3.125 μm (E right).

Article Snippet: Western Blotting (WB) Identification: Exosomes were confirmed by detecting the expression of exosomal markers, including CD63 (Absin, abs132700, CHN), CD9 (Absin, abs136085, CHN), and CD81 (Absin, abs132701, CHN), as well as negative markers, including GM130 (MCE, HY- P82075 , USA) and Calnexin (MCE, HY- P80578 , USA), to assess the purity and quality of the exosomes.

Techniques: Extraction, Transmission Assay, Microscopy, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Antibody-Dependent Cytotoxicity of Monocytes in Preeclampsia Is Associated with Soluble Forms of HLA

doi: 10.3390/ijms262311638

Figure Lengend Snippet: ( a )—CD16+ monocytes obtained one day after delivery from a patient with PE; ( b )—the same monocytes in the placental villus-conditioned culture medium (PVCM) for 3 h; ( c )—the same monocytes in the PVCM and the recombinant histocompatibility antigen HLA-B for 3 h.

Article Snippet: The third well contained PVCM and recombinant human HLA class I histocompatibility antigen, B alpha chain (HLA-B), partial, Code CSB-YP355776HU (Cusabio Biotech, Wuhan, China) at a concentration of 10 μg/mL.

Techniques: Recombinant

Journal: Journal of Biological Chemistry

Article Title: A New Determinant of Endoplasmic Reticulum Localization Is Contained in the Juxtamembrane Region of the Ectodomain of Hepatitis C Virus Glycoprotein E1

doi: 10.1074/jbc.m910400199

Figure Lengend Snippet: FIG. 2. The C-terminal region 99– 192 of HCV E1 protein determines lo- calization in the ER of the reporter CD8. FRT cells stably expressing CD8 (a–b) or CD8-E199–192 (c–f) were analyzed by double indirect immunofluorescence microscopy as detailed under “Experi- mental Procedures” and “Results.” a, c, e, and f, permeabilized cells; b and d, not permeabilized cells. a, c, and e, anti-CD8 mAb OKT8; b and d, polyclonal antibody anti-CD8; f, polyclonal antibody anti- SSRa; a, c, and e, Texas Red-conjugated anti-mouse secondary antibody; b, d, and f, fluorescein-conjugated anti-rabbit sec- ondary antibody.

Article Snippet: The following antibodies were used: mouse mAb OKT8 (anti-CD8 protein), mouse mAb N1 (anti-CD8 protein), rabbit polyclonal anti-CD8 and rabbit polyclonal anti-SSra (16); mouse mAb G10-1 (anti-CD8 protein) (24)2; rabbit polyclonal anti-calnexin (17); rabbit polyclonal anti-calreticulin (Stress-Gene, Canada); rabbit polyclonal anti-alkaline phosphatase (Rockland, Gilbertsville, PA); mouse mAb anti-placental alkaline phosphatase (Chemicon International, Inc., Temecula, CA); peroxidaseconjugated anti-mouse and anti-rabbit IgG (Sigma-Aldrich); Texas Redconjugated anti-mouse IgG and fluorescein-conjugated anti-rabbit IgG (Jackson Immunoresearch Lab, West Grove, PA).

Techniques: Stable Transfection, Expressing, Immunofluorescence, Microscopy

Journal: Journal of Biological Chemistry

Article Title: A New Determinant of Endoplasmic Reticulum Localization Is Contained in the Juxtamembrane Region of the Ectodomain of Hepatitis C Virus Glycoprotein E1

doi: 10.1074/jbc.m910400199

Figure Lengend Snippet: FIG. 6. The juxtamembrane region 99–142 of the ectodomain of HCV E1 protein contains another determi- nant for ER localization. FRT cells sta- bly expressing CD8-E199–142 (a–d) and CD8-SV (e–f) were analyzed by double in- direct immunofluorescence microscopy as detailed under “Experimental Proce- dures” and “Results.” a, c–e, permeabi- lized cells; b and f, non-permeabilized cells. a, c, and e, anti-CD8 OKT8 mAb; b and f, polyclonal antibody anti-CD8; d, polyclonal antibody anti-calnexin; a, c, and e, Texas Red-conjugated anti-mouse secondary antibody; b, d, and f, fluores- cein-conjugated anti-rabbit secondary antibody.

Article Snippet: The following antibodies were used: mouse mAb OKT8 (anti-CD8 protein), mouse mAb N1 (anti-CD8 protein), rabbit polyclonal anti-CD8 and rabbit polyclonal anti-SSra (16); mouse mAb G10-1 (anti-CD8 protein) (24)2; rabbit polyclonal anti-calnexin (17); rabbit polyclonal anti-calreticulin (Stress-Gene, Canada); rabbit polyclonal anti-alkaline phosphatase (Rockland, Gilbertsville, PA); mouse mAb anti-placental alkaline phosphatase (Chemicon International, Inc., Temecula, CA); peroxidaseconjugated anti-mouse and anti-rabbit IgG (Sigma-Aldrich); Texas Redconjugated anti-mouse IgG and fluorescein-conjugated anti-rabbit IgG (Jackson Immunoresearch Lab, West Grove, PA).

Techniques: Expressing, Immunofluorescence, Microscopy

Journal: Journal of Biological Chemistry

Article Title: A New Determinant of Endoplasmic Reticulum Localization Is Contained in the Juxtamembrane Region of the Ectodomain of Hepatitis C Virus Glycoprotein E1

doi: 10.1074/jbc.m910400199

Figure Lengend Snippet: FIG. 5. The TMD region of HCV E1 protein determines localization in the ER of the reporter CD8. FRT cells stably expressing CD8-E1155–192 were analyzed by double indirect immunofluorescence microscopy as detailed under “Experimental Procedures” and in the Results. a, c, and d, permeabilized cells; b, non-permeabilized cells. a and c, anti-CD8 mAb OKT8; b, polyclonal antibody anti-CD8; d, polyclonal antibody anti-calnexin; a and c, Texas red-conjugated anti-mouse secondary antibody; b and d, fluorescein-conjugated anti-rabbit secondary antibody.

Article Snippet: The following antibodies were used: mouse mAb OKT8 (anti-CD8 protein), mouse mAb N1 (anti-CD8 protein), rabbit polyclonal anti-CD8 and rabbit polyclonal anti-SSra (16); mouse mAb G10-1 (anti-CD8 protein) (24)2; rabbit polyclonal anti-calnexin (17); rabbit polyclonal anti-calreticulin (Stress-Gene, Canada); rabbit polyclonal anti-alkaline phosphatase (Rockland, Gilbertsville, PA); mouse mAb anti-placental alkaline phosphatase (Chemicon International, Inc., Temecula, CA); peroxidaseconjugated anti-mouse and anti-rabbit IgG (Sigma-Aldrich); Texas Redconjugated anti-mouse IgG and fluorescein-conjugated anti-rabbit IgG (Jackson Immunoresearch Lab, West Grove, PA).

Techniques: Stable Transfection, Expressing, Immunofluorescence, Microscopy

Journal: Journal of Biological Chemistry

Article Title: A New Determinant of Endoplasmic Reticulum Localization Is Contained in the Juxtamembrane Region of the Ectodomain of Hepatitis C Virus Glycoprotein E1

doi: 10.1074/jbc.m910400199

Figure Lengend Snippet: FIG. 9. CD8-E199–142M protein is ex- pressed on the plasma membrane. HuH-7 cells were transfected with plas- mids expressing CD8-SV (a and b), CD8- E199–142 (c and d), and CD8-E199–142M (e and f) proteins. 36 h post-transfection, the cells were analyzed by single (a and b) and double (c and f) indirect immunoflu- orescence microscopy as detailed under “Experimental Procedures.” a, c, and e, permeabilized cells; b, d, and f, not per- meabilized cells; a–c and e, anti-CD8 mAb OKT8, Texas Red-conjugated anti-mouse secondary antibody; d and f, anti-CD8 polyclonal antibody, fluorescein-conju- gated anti-rabbit secondary antibody.

Article Snippet: The following antibodies were used: mouse mAb OKT8 (anti-CD8 protein), mouse mAb N1 (anti-CD8 protein), rabbit polyclonal anti-CD8 and rabbit polyclonal anti-SSra (16); mouse mAb G10-1 (anti-CD8 protein) (24)2; rabbit polyclonal anti-calnexin (17); rabbit polyclonal anti-calreticulin (Stress-Gene, Canada); rabbit polyclonal anti-alkaline phosphatase (Rockland, Gilbertsville, PA); mouse mAb anti-placental alkaline phosphatase (Chemicon International, Inc., Temecula, CA); peroxidaseconjugated anti-mouse and anti-rabbit IgG (Sigma-Aldrich); Texas Redconjugated anti-mouse IgG and fluorescein-conjugated anti-rabbit IgG (Jackson Immunoresearch Lab, West Grove, PA).

Techniques: Clinical Proteomics, Membrane, Transfection, Expressing, Microscopy

Journal: Journal of Biological Chemistry

Article Title: A New Determinant of Endoplasmic Reticulum Localization Is Contained in the Juxtamembrane Region of the Ectodomain of Hepatitis C Virus Glycoprotein E1

doi: 10.1074/jbc.m910400199

Figure Lengend Snippet: FIG. 12. The juxtamembrane region 99–142 of the ectodomain of HCV E1 determines intracellular localization of the AP protein. HuH-7 cells were transfected with plasmids expressing AP8 (a and b), AP8-SV (c and d), AP8- E199–142 (e and f), and AP8-E199–142M (g and h) proteins. 36 h post-transfection, the cells were analyzed by double indirect immunofluorescence microscopy as de- tailed under “Experimental Procedures.” a, c, e, and g, permeabilized cells, anti-AP mAb, Texas red-conjugated anti-mouse secondary antibody; b, d, f, and h, not permeabilized cells, anti-AP polyclonal antibody, fluorescein-conjugated anti- rabbit secondary antibody.

Article Snippet: The following antibodies were used: mouse mAb OKT8 (anti-CD8 protein), mouse mAb N1 (anti-CD8 protein), rabbit polyclonal anti-CD8 and rabbit polyclonal anti-SSra (16); mouse mAb G10-1 (anti-CD8 protein) (24)2; rabbit polyclonal anti-calnexin (17); rabbit polyclonal anti-calreticulin (Stress-Gene, Canada); rabbit polyclonal anti-alkaline phosphatase (Rockland, Gilbertsville, PA); mouse mAb anti-placental alkaline phosphatase (Chemicon International, Inc., Temecula, CA); peroxidaseconjugated anti-mouse and anti-rabbit IgG (Sigma-Aldrich); Texas Redconjugated anti-mouse IgG and fluorescein-conjugated anti-rabbit IgG (Jackson Immunoresearch Lab, West Grove, PA).

Techniques: Transfection, Expressing, Immunofluorescence, Microscopy

Journal: Nature Neuroscience

Article Title: Cerebrovascular vulnerability and fibrosis in human brain aneurysms

doi: 10.1038/s41593-026-02326-9

Figure Lengend Snippet: a , Heatmap with hierarchical clustering showing cell-to-cell differential interaction strength in brain aneurysms when compared to control cerebrovasculature. y -axis, source; x -axis, target. Red, increased interaction strength in aneurysms; blue, decreased interaction strength in aneurysms. EC, endothelial cell; SMC, smooth muscle cell; FbM, fibromyocyte; FB, fibroblast; aFB, activated fibroblast; mFB, myofibroblast; TC, T-cell; NK, natural killer cell; BC, B-cell; pDC, plasmacytoid dendritic cell; cDC, conventional dendritic cell; pvMϕ, perivascular macrophage; Mo, monocyte; aMϕ, APC5 + macrophage; MG, microglia; AC, astrocyte; Neu, neuron; OL, oligodendrocyte; OPC, oligodendrocyte precursor cell. b , Circle plot of differential interaction strength between activated perivascular fibroblasts (aFB) and myeloid cell populations in aneurysms. Red, increased interaction strength in aneurysms; blue, decreased interaction strength in aneurysms. Line thickness proportional to interaction strength. c , Cell-to-cell communication pathways across cell populations ranked on their differences of overall information flow within inferred networks in aneurysms (red) and control (gray). Red text, pathways statistically over-represented in brain aneurysms; gray text, pathways statistically over-represented in controls; black text, not significant. d , Chord plot showing outgoing cell communication pathways originating from myeloid cells to activated perivascular fibroblasts in aneurysms. Arrow thickness is proportional to interaction strength. e , Scatter-plot of pathways ranked by differential incoming ( y -axis) and outgoing ( x -axis) interaction strength in aFB in brain aneurysms relative to controls. Light blue, aneurysm-specific. Bold, top-nominated outgoing pathway emerging from aFB. f , Representative confocal microscopy analysis of immunostaining CD68 + (magenta) macrophages showing colocalization with CD74 receptor (yellow) and MIF ligand (cyan) in unruptured brain aneurysm (top) and control middle cerebral artery (bottom). DAPI (blue) labels cell nuclei. Colocalization, white. Scale bar, 50 µm. g , Representative confocal microscopy analysis of CD68 + macrophages (magenta) and MIF–CD74 proximity interaction (yellow) in unruptured brain aneurysm (top) and control middle cerebral artery (bottom). DAPI (blue) labels cell nuclei. Colocalization, white. Scale bar, 50 µm.

Article Snippet: Next, we added anti-CD74 antibody (MedChemExpress) at concentration of 5 μg ml −1 and incubated for 30 min followed by washing with medium.

Techniques: Control, Confocal Microscopy, Immunostaining

Journal: Nature Neuroscience

Article Title: Cerebrovascular vulnerability and fibrosis in human brain aneurysms

doi: 10.1038/s41593-026-02326-9

Figure Lengend Snippet: a , Violin plot of the established vascular destabilizing genes MMP9 and SPP1 across cell populations. EC, endothelial cell; SMC, smooth muscle cell; FbM, fibromyocyte; FB, fibroblast; aFB, activated fibroblast; mFB, myofibroblast; TC, T-cell; NK, natural killer cell; BC, B-cell; pDC, plasmacytoid dendritic cell; cDC, conventional dendritic cell; pvMϕ, perivascular macrophage; Mo, monocyte; aMϕ, APC5 + macrophage; MG, microglia; AC, astrocyte; Neu, neuron; OL, oligodendrocyte; OPC, oligodendrocyte precursor cell. b , Enriched biological processes from marker genes in ACP5 + macrophages ordered by statistical significance Benjamini-Hochberg adjusted P < 0.05; dashed line, Benjamini-Hochberg adjusted P cutoff of 0.05. c , Bar graph showing quantitative polymerase chain reaction (qPCR) analysis of MMP9 (left) and ACP5 (right) expression in primary human macrophages treated with interferon (IFN), macrophage inhibitory factor (MIF), and CD74 neutralizing antibody (anti-CD74). Mean ± s.e.m. MMP9: Media vs IFN ( P = 0.2479, ns); IFN vs IFN + MIF ( P = 0.0047, **); IFN + MIF vs IFN+anti-CD74 ( P = 0.0046, **); IFN + MIF vs IFN + MIF+anti-CD74 ( P = 0.017, *). ACP5: Media vs IFN ( P = 0.8114, ns); IFN vs IFN + MIF ( P = 0.0052, **); IFN + MIF vs IFN+anti-CD74 ( P = 0.009, **); IFN + MIF vs IFN + MIF+anti-CD74 ( P = 0.009, **). One-way ANOVA with Tukey’s multiple comparisons test (n = 3 samples). d . Boxplot of the distance (µm) from each aMϕ to its nearest neighbor of each listed cell type (n = 770 aMϕ). Dashed line marks 100 µm. Center lines indicate medians; box limits indicate the 25th and 75th percentiles; whiskers extend to the most extreme data points no more than 1.5 × IQR from the box; points beyond the whiskers are shown as outliers.

Article Snippet: Next, we added anti-CD74 antibody (MedChemExpress) at concentration of 5 μg ml −1 and incubated for 30 min followed by washing with medium.

Techniques: Marker, Real-time Polymerase Chain Reaction, Expressing