Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: GSEA of key genes. (A) The significantly enriched pathways in the HSPG2 high-expression group. (B) The significantly enriched pathways in the BASP1 low-expression group. (C) The significantly enriched pathways in the DSE high-expression group. (D) The significantly enriched pathways in the NRP2 high-expression group.

Article Snippet: The Input group was not added with antibodies and was stored at −20 °C; the IgG group was added with rabbit anti-IgG antibody (Abcam, ab172730, UK); the IP group was added with the target antibodies Anti-FAK antibody(Proteintech, 12,636-1-AP, Rabbit, China) and Anti-NRP2 antibody (Proteintech, 27,193-1-AP, Rabbit, China), and incubated at 4 °C for overnight rotation to form complexes between the antibodies and the target proteins.

Techniques: Expressing

Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: NRP2 mediates the FAK signaling pathway to regulate the malignant progression of GBM cells. (A): NRP2 expression levels in patients from the GSE4290 and GSE68848 data sets. (B): The knockdown effect of sh-NRP2 in U251 and T98G cells was detected by qRT-PCR. (C): WB was used to detect the expressions of pY297-FAK and FAK. (D): The CCK-8 method was used to detect the cell viability of GBM cells in different treatment groups (sh-NC, sh-NRP2, sh-NRP2 + adhesamine). Adhesamine is a FAK activator. (E): The colony formation experiment was used to detect the number of colony formations of GBM cells in different treatment groups; (F): The scratch healing experiment was used to detect the migration ability of GBM cells in different treatment groups; (G): The Transwell experiment was used to detect the invasion ability of GBM cells in different treatment groups; (H): The Annexin V/PI double staining method was used to detect the apoptosis level of cells in different treatment groups. (I): Western blot was used to detect the protein expression of pro-apoptotic indicators clever-caspase3, clever-PARP, BAX and pro-apoptotic indicator BCL2 in different treatment groups. * indicates P < 0.05, **** indicates P < 0.0001.

Article Snippet: The Input group was not added with antibodies and was stored at −20 °C; the IgG group was added with rabbit anti-IgG antibody (Abcam, ab172730, UK); the IP group was added with the target antibodies Anti-FAK antibody(Proteintech, 12,636-1-AP, Rabbit, China) and Anti-NRP2 antibody (Proteintech, 27,193-1-AP, Rabbit, China), and incubated at 4 °C for overnight rotation to form complexes between the antibodies and the target proteins.

Techniques: Expressing, Knockdown, Quantitative RT-PCR, CCK-8 Assay, Migration, Double Staining, Western Blot

Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: NRP2 affects the Focal-adhesion pathway through FAK to regulate the progression of GBM. (A): Western blot was used to detect the expression levels of FAK and its phosphorylated proteins in GBM cells of different treatment groups (sh-NC, sh-NRP2). (B): Immunofluorescence co-localization was used to verify the binding of NRP2 and FAK. (C): Immunoprecipitation was used to verify the binding of NRP2 and FAK in different groups.

Article Snippet: The Input group was not added with antibodies and was stored at −20 °C; the IgG group was added with rabbit anti-IgG antibody (Abcam, ab172730, UK); the IP group was added with the target antibodies Anti-FAK antibody(Proteintech, 12,636-1-AP, Rabbit, China) and Anti-NRP2 antibody (Proteintech, 27,193-1-AP, Rabbit, China), and incubated at 4 °C for overnight rotation to form complexes between the antibodies and the target proteins.

Techniques: Western Blot, Expressing, Immunofluorescence, Binding Assay, Immunoprecipitation

Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: NRP2 promotes the malignant progression of GBM by activating FAK. U251 and T98G cell groups: sh-NC, sh-NRP2, sh-NRP2 + oe-FAK. (A): qRT-PCR was used to detect the mRNA level of FAK; (B): WB was used to detect the protein expression of pY397-FAK and FAK; (C): CCK-8 was used to detect cell viability; (D): Scratch test was used to detect the migration ability of cells; (E): Transwell test was used to detect the invasion ability of cells; (F): WB was used to detect the protein expression of apoptotic proteins clever-caspase3, clever-PARP, BAX and anti-apoptotic protein BCL2. * indicates P < 0.05.

Article Snippet: The Input group was not added with antibodies and was stored at −20 °C; the IgG group was added with rabbit anti-IgG antibody (Abcam, ab172730, UK); the IP group was added with the target antibodies Anti-FAK antibody(Proteintech, 12,636-1-AP, Rabbit, China) and Anti-NRP2 antibody (Proteintech, 27,193-1-AP, Rabbit, China), and incubated at 4 °C for overnight rotation to form complexes between the antibodies and the target proteins.

Techniques: Quantitative RT-PCR, Expressing, CCK-8 Assay, Migration

Journal: Translational Oncology

Article Title: The brain imaging feature-related gene NRP2 drives the malignant progression of glioblastoma through the FAK pathway: a Mendelian randomization study

doi: 10.1016/j.tranon.2026.102732

Figure Lengend Snippet: The nude mouse experiment demonstrates that NRP2 mediates the Focal-adhesion pathway to regulate the formation of GBM tumors. (A): Nude mice were subcutaneously injected with sh-NRP2 and sh-NC stably transfected T98G cells. Comparison of tumor size in sh-NRP2 nude mice treated with FAK activator adhesamine. (B): Tumor weights of different treatment groups of nude mice. (C): Tumor volumes of different treatment groups of nude mice. (D): IHC detection of the expression of Ki67, BCL2 and BAX in tumor tissues; (E): WB detection of the expression of pY297-FAK and FAK in tumor tissues.

Article Snippet: The Input group was not added with antibodies and was stored at −20 °C; the IgG group was added with rabbit anti-IgG antibody (Abcam, ab172730, UK); the IP group was added with the target antibodies Anti-FAK antibody(Proteintech, 12,636-1-AP, Rabbit, China) and Anti-NRP2 antibody (Proteintech, 27,193-1-AP, Rabbit, China), and incubated at 4 °C for overnight rotation to form complexes between the antibodies and the target proteins.

Techniques: Injection, Stable Transfection, Transfection, Comparison, Expressing

Journal: bioRxiv

Article Title: A Csf1r lineage gives rise to dermal lymphatic endothelial cells

doi: 10.64898/2026.03.17.712362

Figure Lengend Snippet: A,B Strategy for the combined Csf1r-iCre -mediated lineage tracing and targeting of Prox1 ( A ) and representative immunofluorescence staining with the indicated markers of E15.5 dermis from a heterozygously targeted Csf1r-iCre ; Prox1 fl(Egfp)/+ mouse ( B ) (scale bars: 25 μm). C,D Representative images of E15.5 Prox1 fl/fl embryos with or without Csf1r-iCre (scale bars: 1 mm) ( C ) and table showing the frequency of embryos displaying the indicated phenotype ( D ); n = 20 Prox1 fl/+ , n = 22 Prox1 fl/fl , n = 22 Csf1r-iCre ; Prox1 fl/+ , n = 19 Csf1r-iCre ; Prox1 fl/fl from 11 litters. E Immunofluorescence staining with the indicated markers of E15.5 dermis of Csf1r-iCre ; Rosa tdTom ; Prox1 fl(Egfp)/+ and Csf1r-iCre ; Rosa tdTom ; Prox1 fl(Egfp)/fl(Egfp) littermate mice identifies erythrocytes in mutant dermal lymphatic vessels that are co-labelled for TOM and GFP (scale bars: 100 μm). F Representative immunofluorescence staining with the indicated markers of E15.5 Prox1 fl(Egfp)/fl(Egfp) (no Cre, normal PROX1 function), Csf1r-iCre ; Prox1 fl(Egfp)/+ (heterozygous PROX1 deficiency) and E15.5 Csf1r-iCre ; Prox1 fl(Egfp)/fl(Egfp) (homozygous PROX1 deficiency) dermis illustrates that TER119+ erythrocytes are located in NRP2+ lymphatic vessels of PROX1-deficient embryos. The square indicates an area shown at higher magnification in the adjacent panels and shown for the different markers also in grey scale (Scale bars: 100 μm). Arrows indicate TER119+ erythrocytes in lymphatic vessels.

Article Snippet: Embryonic dorsal dermis was dissected from formaldehyde-fixed embryos and then incubated in PBS containing 2% serum-free protein block (DAKO), 2% bovine serum albumin and 0.4% Triton X-100 before staining with a combination of the following primary antibodies: goat anti-mouse NRP2 (R&D Systems #AF567, 1:100), rabbit anti-mouse PROX1 (Biolegend # 925202, 1:50), goat anti-mouse PROX1 (R&D Systems # AF2727, 1:100), rabbit anti-mouse LYVE1 (Angiobio # 11-034, 1:100), rat anti-mouse PECAM1 (BD Pharmigen # 553370, 1:50), rat anti-mouse EMCN (Santacruz # sc-65495, 1:50), goat anti-mouse FLT4 (R&D Systems # AF743, 1:100), rat anti-mouse TER119 (Biolegend # 116241, 1:100), rabbit anti-mouse GFP (MBL # 598, 1:200), goat anti-mouse TOM (Origene #AB1140-100, 1:250), or rat anti-mouse TOM (Chromotek #5F8, 1:200).

Techniques: Immunofluorescence, Staining, Mutagenesis

Journal: Journal of Virology

Article Title: Differential expression of viral entry protein neuropilin 1 (NRP1) and neuropilin 2 (NRP2) in fatal COVID-19

doi: 10.1128/jvi.01384-25

Figure Lengend Snippet: NRP1 is expressed on endothelial and mononuclear cells in fatal COVID-19 cardiac autopsy tissue. ( A ) CODEX image with DAPI displayed in blue, NRP1 in red and CD31 in yellow. Co-localization of NRP1 and the endothelial marker CD31/PECAM1 (indicated by arrows). ( B ) Capillaries (arrows) and larger vessels (inset; arrowheads) expressed NRP1 by IHC. ( C ) H&E staining corresponding to B. ( D ) NRP1 expression in larger vessels and small capillaries (arrowheads) and mononuclear cells (arrows). Corresponding H&E stain in inset. ( E ) CD68-positive macrophages lining the vascular bed of a larger vessel (arrows). Adipocytes are marked by asterisks (*). A–C , patient 2; D–E , patient 12 . Magnification 400× for B–E . Scale bars correspond to 50 µm. ( F ) Automatic cell classification identified 19-cell subsets across compartments in re-analyzed data set of fatal COVID-19 hearts . No annotated cluster for adipocytes and erythroid cells. Gene expression along clusters for ACE2 ( G ), NRP1 ( H ), and NRP2 ( I ). ( K ) Dot plot summarizing the expression levels with dot size indicating the percentage of cells expressing the relevant gene and saturation corresponding to average expression level. All plots were generated using the Scanpy package in Python.

Article Snippet: HEK293 cells stably expressing NRP2v2 (RC220706, Origene Technologies) were generated accordingly and checked for expression by Western blot of cell lysates using anti-NRP2 antibody (AF2215, R&D Systems, Abingdon, UK).

Techniques: Marker, Staining, Expressing, Gene Expression, Generated

Journal: Journal of Virology

Article Title: Differential expression of viral entry protein neuropilin 1 (NRP1) and neuropilin 2 (NRP2) in fatal COVID-19

doi: 10.1128/jvi.01384-25

Figure Lengend Snippet: Alveolar and syncytial macrophages express NRP1 and NRP2 in fatal COVID-19 lung autopsy tissue. ( A ) CODEX image showing syncytial macrophages co-expressing CD68 and NRP1 (arrows, insets). ( B, C ) IHC of the same lung vessel with corresponding H&E inset. ( B ) NRP1 protein in alveolar macrophages (arrows) and blood mononuclear cells (black arrowhead). Upper inset: NRP1 expression in vascular endothelium (white arrowhead). Lower inset: H&E stain of corresponding region. ( C ) IHC of NRP2 expression in a syncytial macrophage (arrow) in serial section of B. IHC of NRP1 ( D ) and CD68 ( E ) positive syncytial macrophage in the lung (arrows). ( F–H ) CODEX images highlighting macrophages in the lung with co-expression of IBA1 ( F ), CD68 ( G ), and NRP1 ( H ). Arrowheads indicate macrophages with ruffled membrane segments. A–C and F–H , patient 3; D–E , patient 20 . Scale bars correspond to 50 µm unless stated otherwise. CODEX color code: nuclei (DAPI, blue), pancytokeratin (panCK, white), NRP1 (red), IBA1 (magenta), CD68 (yellow). ( I ) Automatic cell classification identified 28-cell subsets across compartments in re-analyzed data set of fatal COVID-19 lungs . Gene expression along clusters for ACE2 ( K ), NRP1 ( L ), and NRP2 ( M ). ( N ) UMAP embedding showing SARS-CoV-2 RNA + cells (black dots). ( O ) UMAP plot showing co-expression of acknowledged SARS-CoV-2 entry factors ACE2 and TMPRSS2 . SARS-CoV-2 RNA ( N ) was detected in cells devoid of ACE2 and TMPRSS2 expression. ( P ) Dot plot summarizing expression of genes associated with SARS-CoV-2 and neuropilin pathways. FLT1 , VEGF receptor 1; KDR , VEGF receptor 2; FLT4 , VEGF receptor 3. All plots were generated using the scanpy package in Python.

Article Snippet: HEK293 cells stably expressing NRP2v2 (RC220706, Origene Technologies) were generated accordingly and checked for expression by Western blot of cell lysates using anti-NRP2 antibody (AF2215, R&D Systems, Abingdon, UK).

Techniques: Expressing, Staining, Membrane, Gene Expression, Generated

Journal: Journal of Virology

Article Title: Differential expression of viral entry protein neuropilin 1 (NRP1) and neuropilin 2 (NRP2) in fatal COVID-19

doi: 10.1128/jvi.01384-25

Figure Lengend Snippet: NRP1 expression on lymphocytes may enable SARS-CoV-2 infection in the spleen and lymph node of fatal COVID-19. ( A ) CODEX image of a lymph follicle in the spleen in fatal COVID-19. A subset of B lymphocytes co-expressed NRP1, while cytotoxic T cells were NRP1 negative. The inset shows that the depicted lymph follicle is largely composed of CD20-positive B cells. ( B ) Corresponding region in NRP1 IHC revealed strong NRP1 expression in follicular lymphocytes, while the same region was negative for NRP2 ( C ). ( D ) H&E stain of corresponding serial section. ( E ) CODEX image of a lymph node from a fatal COVID-19 case. CD8 and CD4 T cells did not co-express NRP1, while pDCs in lymph node, identified by the expression of CD123, co-expressed NRP1 ( F ). Scale bars correspond to 50 µm unless stated otherwise. A–D , COVID-19 patient 2; E–F , COVID-19 patient 3 . CODEX color code: nuclei (DAPI, blue), NRP1 (red), B lymphocyte marker CD20 (yellow), cytotoxic T-cell marker CD8 (cyan), helper T-cell marker CD4 (green) and CD123 (white). ( G ) RNA scope analysis of spleen and lymph node from fatal COVID-19 cases (patient 2 and 3; ) and influenza-related death (patient 5; ) with corresponding areas in H&E (left column), using serial sections. In COVID-19 cases, lymphocytes within the spleen and lymph nodes displayed positive RNAscope signals for SARS-CoV-2 RNA. In contrast, lymph node tissue from the influenza patient was largely negative. Bottom row: RNAscope probes with defined targets were applied to influenza lymph node tissue (patient 5), including a positive control probe set (3-plex targeting POLR2A , PPIB , and UBC ) and a negative control probe (targeting the dapB gene of Bacillus subtilis ). Scale bars are provided per column. FISH images in the far-right column show a twofold magnification of the middle column with areas of interest highlighted in white boxes.

Article Snippet: HEK293 cells stably expressing NRP2v2 (RC220706, Origene Technologies) were generated accordingly and checked for expression by Western blot of cell lysates using anti-NRP2 antibody (AF2215, R&D Systems, Abingdon, UK).

Techniques: Expressing, Infection, Staining, Marker, RNAscope, Positive Control, Negative Control

Journal: Journal of Virology

Article Title: Differential expression of viral entry protein neuropilin 1 (NRP1) and neuropilin 2 (NRP2) in fatal COVID-19

doi: 10.1128/jvi.01384-25

Figure Lengend Snippet: Mast cells strongly express NRP2. ( A ) H&E staining of lung tissue from a fatal COVID-19 case. ( B , C ) Corresponding region in CODEX. ( D ) CD8+ T-cell infiltrate in surrounding area. ( E ) NRP2-positive cells (arrows). ( F ) Mast cell tryptase identified NRP2-positive cells as mast cells. ( G ) A subset of mast cells also expressed NRP1 (white arrows). NRP1-negative mast cells are marked with black arrows. ( H ) Mast cells were also positive for CD16. A–H , patient 2 . Scale bars correspond to 50 µm. CODEX color code: DAPI (blue), CD8 (cyan), NRP2 (green), mast cell tryptase (yellow), NRP1 (red), and CD16 (magenta).

Article Snippet: HEK293 cells stably expressing NRP2v2 (RC220706, Origene Technologies) were generated accordingly and checked for expression by Western blot of cell lysates using anti-NRP2 antibody (AF2215, R&D Systems, Abingdon, UK).

Techniques: Staining

Journal: Journal of Virology

Article Title: Differential expression of viral entry protein neuropilin 1 (NRP1) and neuropilin 2 (NRP2) in fatal COVID-19

doi: 10.1128/jvi.01384-25

Figure Lengend Snippet: Binding of soluble S1′ spike fragment requires presence of NRP2. HEK293 cells endogenously expressing NRP1 bound soluble S1 spike protein ( A–C ) but not soluble S1′ ( E–G ). HEK293 cells stably transfected to express NRP2 bound soluble S1 ( I–L ) and also S1′ ( N–P ). DAPI nuclear staining ( A, E, I, N ). Anti-HA IF for S1 or S1′ ( B, F, K, O ); merged DAPI/IF ( C, G, L, P ); isotype controls ( D, H, M, Q ).

Article Snippet: HEK293 cells stably expressing NRP2v2 (RC220706, Origene Technologies) were generated accordingly and checked for expression by Western blot of cell lysates using anti-NRP2 antibody (AF2215, R&D Systems, Abingdon, UK).

Techniques: Binding Assay, Expressing, Stable Transfection, Transfection, Staining