Journal: The Journal of Biological Chemistry

Article Title: Matriptase-2-mediated suppression of hepatic hepcidin expression in mice requires hepatocyte neogenin

doi: 10.1016/j.jbc.2026.111142

Figure Lengend Snippet: Mt2 expression markedly reduces Neo1 levels in Hep3B cells . A , diagram of Mt2 and fMt2. The MYC (m) and FLAG (f) epitopes were added to the C-terminus of Mt2. B , diagram of fNeo1. The MYC (m) and FLAG (f) epitopes were added to the C-terminus of Neo1. C , coexpression of fMt2 with fNeo1 markedly reduces fNeo1 levels in whole cell extracts (input) and on cell surface by biotinylation. AP, aprotinin. Experiments were repeated three times with consistent results. D , quantification of cell surface fNeo1 bands in C ( panel 4 ). The relative amounts to Na + K + ATPase ( panel-5 ) are presented (n = 3). The data shown are means ± SD. One-way ANOVA was used to analyze the data. ns, no statistical difference. ∗, p < 0.05; ∗∗∗∗, p < 0.0001. NEO1, neogenin.

Article Snippet: We purchased murine Neo1 ORF ( NM_008684 ) with a C-terminal FLAG/MYC epitope (fNeo1) in pCMV6 vector (#MR226235) and murine Mt2 ORF ( NM_027902.1 ) with a C-terminal FLAG/MYC epitope (fMt2) in pCMV6 vector (#MR210781) from OriGene Technologies Inc.

Techniques: Expressing

Journal: The Journal of Biological Chemistry

Article Title: Matriptase-2-mediated suppression of hepatic hepcidin expression in mice requires hepatocyte neogenin

doi: 10.1016/j.jbc.2026.111142

Figure Lengend Snippet: Hepcidin is an iron regulatory hormone that is secreted mainly by hepatocytes . A , hepcidin inhibits iron efflux from duodenum, spleen, and the liver into the circulation by blocking the plasma-membrane iron exporter, ferroportin. B , diagram of the key components that are involved in the induction of hepcidin expression in the liver. C , cartoon of NEO1 protein. It contains four immunoglobulin-like (Ig) domain, six fibronectin III domains, a transmembrane domain, and a cytoplasmic domain (CD). D , diagram of MT2 protein. Cyto: cytoplasmic domain. transmembranedomain. SEA: sea urchin sperm protein, enteropeptidase agrin. CUB: complement protein subcomponents C1r/C1s, urchin embryonic growth factor and bone morphogenetic protein one domain. L: low-density lipoprotein receptor class-A domain. Catalytic: serine protease (S/P) catalytic domain. The arrow indicates the predicted autocleavage activation site. E , iron induction of hepcidin expression by increases in Bmp6 gene transcription and TfR2 protein stabilization. F , a proposed model for the function of MT2. NEO1, neogenin.

Article Snippet: We purchased murine Neo1 ORF ( NM_008684 ) with a C-terminal FLAG/MYC epitope (fNeo1) in pCMV6 vector (#MR226235) and murine Mt2 ORF ( NM_027902.1 ) with a C-terminal FLAG/MYC epitope (fMt2) in pCMV6 vector (#MR210781) from OriGene Technologies Inc.

Techniques: Blocking Assay, Clinical Proteomics, Membrane, Expressing, Activation Assay

Journal: The Journal of Biological Chemistry

Article Title: Matriptase-2-mediated suppression of hepatic hepcidin expression in mice requires hepatocyte neogenin

doi: 10.1016/j.jbc.2026.111142

Figure Lengend Snippet: Expression of exogenously administered Mt2 in the liver of Tmprss6 −/− mice increases Neo1 levels . A , diagram of Mt2, fMt2, and fMt2 mask . B , experimental design to determine the effects of exogenously administered Mt2 on Neo1 levels. Eight-week-old Tmprss6 −/− littermates on a mixed B6/129 background were intraperitoneally injected with AAV8-fMt2 vectors at ∼8 x 10 11 or ∼4 x 10 12 viral genome-particles per mouse or AAV8-fMt2 mask viral vectors at ∼4 x 10 12 viral genome-particles per mouse. Injection of sterilized PBS vehicle was included as a control. Age and gender-matched WT littermates were included as additional controls. Mice were euthanized for analysis 3 weeks after injection. All mice were fed a standard diet containing 240-ppm iron. Each group consists of at least five mice with similar numbers of male and female. C , qRT-PCR analysis of Tmprss6 mRNA in the liver. Results are expressed as the amount relative to that of β-actin for each sample. The mean values and SD are presented. D , representative images of Western blot analysis for Neo1, the FLAG epitope of fMt2, Tfr2, and β-actin in the liver membrane preparations. E , qRT-PCR analysis of Hamp mRNA in the liver. F , serum iron assay. G ,. quantification of Neo1 bands in D . H , quantification of Tfr2 bands in D . The relative amounts to β-actin are presented. One-way ANOVA was used to analyze the data relative to WT mice. ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. I , models for Neo1/Hjv induction of hepcidin expression in the absence and presence of Mt2. NEO1, neogenin.

Article Snippet: We purchased murine Neo1 ORF ( NM_008684 ) with a C-terminal FLAG/MYC epitope (fNeo1) in pCMV6 vector (#MR226235) and murine Mt2 ORF ( NM_027902.1 ) with a C-terminal FLAG/MYC epitope (fMt2) in pCMV6 vector (#MR210781) from OriGene Technologies Inc.

Techniques: Expressing, Injection, Control, Quantitative RT-PCR, Western Blot, FLAG-tag, Membrane, Iron Assay

Journal: The Journal of Biological Chemistry

Article Title: Matriptase-2-mediated suppression of hepatic hepcidin expression in mice requires hepatocyte neogenin

doi: 10.1016/j.jbc.2026.111142

Figure Lengend Snippet: Increased Mt2 expression in the liver of WT mice does not reduce Neo1 . Eight-week-old 129S mice were intraperitoneally injected with AAV8-fMt2 vectors at ∼8 x 10 11 , ∼3 x 10 12 , or ∼6 x 10 12 viral genome-particles per mouse or AAV8-fMt2 mask viral vectors at ∼4 x 10 12 viral genome-particles per mouse. Injection of sterilized PBS vehicle was included as a control. Mice were euthanized for analysis 3 weeks after injection. All mice were fed a standard diet containing 240-ppm iron. Each group consists of at least five mice with similar numbers of male and female. A , qRT-PCR analysis of Tmprss6 mRNA in the liver. Results are expressed as the amount relative to that of β-actin for each sample. The mean values and SD are presented. B , representative images of Western blot analysis for Neo1, the FLAG epitope of fMt2, Tfr2, Hjv, and β-actin in the liver membrane preparations. C , qRT-PCR analysis of Hamp mRNA in the liver. D , serum iron assay. E – G , quantification of Neo1, Hjv, and Tfr2 bands in B . The relative amounts to β-actin are presented. One-way ANOVA was used to analyze the data relative to PBS-injected WT controls. ns, no statistical difference. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗∗, p < 0.0001. NEO1, neogenin.

Article Snippet: We purchased murine Neo1 ORF ( NM_008684 ) with a C-terminal FLAG/MYC epitope (fNeo1) in pCMV6 vector (#MR226235) and murine Mt2 ORF ( NM_027902.1 ) with a C-terminal FLAG/MYC epitope (fMt2) in pCMV6 vector (#MR210781) from OriGene Technologies Inc.

Techniques: Expressing, Injection, Control, Quantitative RT-PCR, Western Blot, FLAG-tag, Membrane, Iron Assay

Journal: The Journal of Biological Chemistry

Article Title: Matriptase-2-mediated suppression of hepatic hepcidin expression in mice requires hepatocyte neogenin

doi: 10.1016/j.jbc.2026.111142

Figure Lengend Snippet: Ablation of both Tmprss6 and hepatic Neo1 does not affect iron induction of hepcidin expression . Five-week-old Neo1 fl/fl ; Alb-Cre + ; Tmprss6 −/− mice of both genders were fed an IDD, ICD, or HID for 4 weeks before euthanasia for analysis. These animal models were generated in the same setting of studies as described in the legend to for Neo1 fl/fl ; Alb-Cre - and Neo1 fl/fl ; Alb-Cre + mice. All presented data in this figure are generated from male mice. A , serum iron assay. One-way ANOVA was used for analysis. B , liver nonheme iron assay. Green asterisks represent one-way ANOVA analysis of the data from mice fed the same iron diet relative to the corresponding WT ( Neo1 fl/fl ; Alb-Cre - ; Tmprss6 +/+ ) controls. Red asterisks represent Two-tailed student-T test between Neo1 fl/fl ; Alb-Cre + ; Tmprss6 +/+ and Neo1 fl/fl ; Alb-Cre + ; Tmprss6 −/− mice fed the same iron diet. C , qRT-PCR analysis of Hamp mRNA levels in the liver. All qRT-PCR results are expressed as the amount relative to that of β-actin for each sample. D , serum hepcidin assay. E , qRT-PCR analysis of Id1 mRNA levels in the liver. F or G , normalized hepatic Hamp and Id1 mRNA levels to those of liver nonheme iron (C/B and E/B). Each group consists of at least four animals. The means ± SD are presented. ns, no statistical difference. ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. H , a model for Mt2 suppression of hepcidin expression in hepatocytes. IDD, iron deficient diet; NEO1, neogenin.

Article Snippet: We purchased murine Neo1 ORF ( NM_008684 ) with a C-terminal FLAG/MYC epitope (fNeo1) in pCMV6 vector (#MR226235) and murine Mt2 ORF ( NM_027902.1 ) with a C-terminal FLAG/MYC epitope (fMt2) in pCMV6 vector (#MR210781) from OriGene Technologies Inc.

Techniques: Expressing, Generated, Iron Assay, Two Tailed Test, Quantitative RT-PCR

Journal: The Journal of Headache and Pain

Article Title: Expression of melatonin receptors in trigeminal and sphenopalatine ganglia: potential targets for primary headache disorders

doi: 10.1186/s10194-025-02215-9

Figure Lengend Snippet: Expression of MT1 and MT2 genes in rat TG, SPG and hypothalamus. ( A ) Comparison of MT1 and MT2 mRNA levels in the TG of male and female rats shows that MT1 expression is significantly higher than MT2. ( B ) In both the hypothalamus (HT) and SPG of male rats, MT1 mRNA levels are also significantly higher than MT2. Data are presented as mean ± SEM, statistical significance indicate as *** p < 0.001, ** p < 0.01, * p < 0.05, and n = 6

Article Snippet: MT2 (AMR-032) , 1:200 , Rabbit , Alomone Labs, Jerusalem, Israel , AB_2340995.

Techniques: Expressing, Comparison

Journal: The Journal of Headache and Pain

Article Title: Expression of melatonin receptors in trigeminal and sphenopalatine ganglia: potential targets for primary headache disorders

doi: 10.1186/s10194-025-02215-9

Figure Lengend Snippet: MT1 and MT2 immunoreactivity in TG. ( A ) positive MT1 immunoreactivity was observed in the cytoplasm and nuclei of TG neurons (thick arrows) and in SGCs (arrowheads). ( B ) MT2 immunoreactivity was observed predominantly in the cytoplasm of neurons (thick arrows), in some cases in their nuclei (thick arrows), and in Aδ-fibers (thin arrows). Negative cells are indicated by asterisks in both ( A ) and ( B ). Blue color represents nuclei staining with DAPI. ( C ) Bar graphs show the number of MT1 and MT2 immunoreactive TG neurons in male and female rats. There was no significant difference in MT1 or MT2 protein expression between sexes. However, the number of MT1-immunoreactive cells was approximately twofold higher than that of MT2 in both males and females. Data are presented as the mean ± SEM, n = 6 and * p < 0.05

Article Snippet: MT2 (AMR-032) , 1:200 , Rabbit , Alomone Labs, Jerusalem, Israel , AB_2340995.

Techniques: Staining, Expressing

Journal: The Journal of Headache and Pain

Article Title: Expression of melatonin receptors in trigeminal and sphenopalatine ganglia: potential targets for primary headache disorders

doi: 10.1186/s10194-025-02215-9

Figure Lengend Snippet: Double immunohistochemistry of MT2 with CGRP or RAMP1 in TG. ( A and D ) MT2 immunoreactivity (green fluorescence) was observed in cytoplasm of TG neurons (thick arrows), and in Aδ- fibers (arrowheads). ( B ) CGRP (red fluorescence) was expressed in small to medium-sized TG neurons (thick arrows) as well as in C-fibers (arrowhead). ( C ) MT2 co-localized with CGRP (yellow-orange) in the cytoplasm of small to medium-sized neurons (thick arrows). ( E ) RAMP1 (red fluorescence) was expressed in the cytoplasm of medium to large- sized TG neurons (thick arrow) and in Aδ-fibers (arrowhead). ( F ) The merged imaged showed that MT2 was co-localized (yellow) with RAMP1 in the cytoplasm of medium to large-sized neurons (thick arrow) as well as in the Aδ- fibers (arrowhead)

Article Snippet: MT2 (AMR-032) , 1:200 , Rabbit , Alomone Labs, Jerusalem, Israel , AB_2340995.

Techniques: Immunohistochemistry, Fluorescence

Journal: The Journal of Headache and Pain

Article Title: Expression of melatonin receptors in trigeminal and sphenopalatine ganglia: potential targets for primary headache disorders

doi: 10.1186/s10194-025-02215-9

Figure Lengend Snippet: Double immunohistochemistry of MT2 and CASPR in trigeminal Aδ-fibers. ( A ) MT2 receptors (green) were found in the Aδ-fibers. ( B ) CASPR immunoreactivity (red) was used to label the paranodal regions of nodes of Ranvier. ( C ) MT2 and CASPR were co-expressed in Aδ-fibers. The blue color represents nuclei staining with DAPI

Article Snippet: MT2 (AMR-032) , 1:200 , Rabbit , Alomone Labs, Jerusalem, Israel , AB_2340995.

Techniques: Immunohistochemistry, Staining

Journal: The Journal of Headache and Pain

Article Title: Expression of melatonin receptors in trigeminal and sphenopalatine ganglia: potential targets for primary headache disorders

doi: 10.1186/s10194-025-02215-9

Figure Lengend Snippet: Double immunohistochemistry of MT2 with CASPR and CGRP in rat dura mater. ( A and D ) Thin MT2-immunoreactive axons (green, arrows) were observed in the rat dura mater. ( B ) These fibers were identified as Aδ-fibers based on CASPR immunoreactivity (red) showing the characteristic bowtie-shaped paranodal pattern (arrowheads). ( A - C ) Fiber bundles were often seen flanking dural blood vessels (asterisks). ( C ) MT2 and CASPR were observed to be co-expressed in the same Aδ-fibers. ( E ) CGRP immunoreactivity (red, arrowheads) was detected in c-fibers throughout the rat dura mater in a granular, pearl-like pattern. ( F ) Merged images show intertwined C- and Aδ-fibers, expressing CGRP and MT2 respectively, projecting throughout the dura mater

Article Snippet: MT2 (AMR-032) , 1:200 , Rabbit , Alomone Labs, Jerusalem, Israel , AB_2340995.

Techniques: Immunohistochemistry, Expressing

Journal: The Journal of Headache and Pain

Article Title: Expression of melatonin receptors in trigeminal and sphenopalatine ganglia: potential targets for primary headache disorders

doi: 10.1186/s10194-025-02215-9

Figure Lengend Snippet: Localization of MT2 immunoreactivity in the SPG and its co-localization with VIP and PACAP. ( A , D and G ) MT2 immunoreactivity (green) was observed in the cytoplasm of neurons (thick arrows), and in fibers (arrowhead). ( B and E ) VIP immunoreactivity (red) was detected in the cytoplasm of many medium-sized neurons (thick arrows) and in a few fibers (arrowheads). ( C and F ) MT2 was co-localized with VIP in the cytoplasm of medium sized neurons (yellow-orange, thick arrows). D , E and F are higher-magnification views of images A , B and C , respectively. ( H ) PACAP immunoreactivity (red) was found in the cytoplasm of SPG neurons (thick arrows). ( I ) Double staining of MT2 and PACAP showed that MT2 was co-localized with PACAP in the cytoplasm of SPG neurons (yellow-orange, thick arrow)

Article Snippet: MT2 (AMR-032) , 1:200 , Rabbit , Alomone Labs, Jerusalem, Israel , AB_2340995.

Techniques: Double Staining

Journal: The Journal of Headache and Pain

Article Title: Expression of melatonin receptors in trigeminal and sphenopalatine ganglia: potential targets for primary headache disorders

doi: 10.1186/s10194-025-02215-9

Figure Lengend Snippet: MT1 and MT2 immunoreactivity in the DRG. ( A ) MT1 expression in DRG was observed in both the cytoplasm and nuclei of neurons (thick arrow) and in SGCs (arrowheads). ( B ) MT2 immunoreactivity was localized in the cytoplasm of neurons and in some cases in their nuclei (thick arrow), and in the Aδ-fibers (thin arrows). The blue color represents nuclei staining with DAPI

Article Snippet: MT2 (AMR-032) , 1:200 , Rabbit , Alomone Labs, Jerusalem, Israel , AB_2340995.

Techniques: Expressing, Staining