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Cusabio fcn3
Low <t>FCN3</t> expression in HCC affected Treg cell activation. ( A ) Kaplan-Meier survival analysis showing the correlation between FCN3 levels and overall survival of HCC patients. ( B - D ) qRT-PCR, WB and IHC analysis of FCN3 expression in tumor ( T ) and adjacent non-tumor ( N ) tissues from HCC patients. ( E - F ) The levels of FCN3 in normal hepatocyte THLE-2 and HCC cell lines (HepG2, Hep3B, HCC-LM3) were assessed by qRT-PCR and WB. ( G - H ) qRT-PCR and WB analysis demonstrating the mRNA and protein levels of Treg cell markers FOXP3 and CD25 in clinical samples. ( I ) <t>ELISA</t> quantification of TGF-β1 and IL-10 in HCC tumor and adjacent non-tumor tissues. ( J - K ) The levels of FOXP3 and CD25 in PBMCs co-cultured with FCN3-knockdown or FCN3-overexpressing HCC cells were evaluated by qRT-PCR and WB. ( L ) Flow cytometry analysis of CD4⁺CD25⁺FOXP3 + Treg cell proportion in PBMCs within the co-culture systems. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC/ OE-NC
Fcn3, supplied by Cusabio, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin"

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

Journal: Cellular Oncology (Dordrecht, Netherlands)

doi: 10.1007/s13402-025-01159-1

Low FCN3 expression in HCC affected Treg cell activation. ( A ) Kaplan-Meier survival analysis showing the correlation between FCN3 levels and overall survival of HCC patients. ( B - D ) qRT-PCR, WB and IHC analysis of FCN3 expression in tumor ( T ) and adjacent non-tumor ( N ) tissues from HCC patients. ( E - F ) The levels of FCN3 in normal hepatocyte THLE-2 and HCC cell lines (HepG2, Hep3B, HCC-LM3) were assessed by qRT-PCR and WB. ( G - H ) qRT-PCR and WB analysis demonstrating the mRNA and protein levels of Treg cell markers FOXP3 and CD25 in clinical samples. ( I ) ELISA quantification of TGF-β1 and IL-10 in HCC tumor and adjacent non-tumor tissues. ( J - K ) The levels of FOXP3 and CD25 in PBMCs co-cultured with FCN3-knockdown or FCN3-overexpressing HCC cells were evaluated by qRT-PCR and WB. ( L ) Flow cytometry analysis of CD4⁺CD25⁺FOXP3 + Treg cell proportion in PBMCs within the co-culture systems. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC/ OE-NC
Figure Legend Snippet: Low FCN3 expression in HCC affected Treg cell activation. ( A ) Kaplan-Meier survival analysis showing the correlation between FCN3 levels and overall survival of HCC patients. ( B - D ) qRT-PCR, WB and IHC analysis of FCN3 expression in tumor ( T ) and adjacent non-tumor ( N ) tissues from HCC patients. ( E - F ) The levels of FCN3 in normal hepatocyte THLE-2 and HCC cell lines (HepG2, Hep3B, HCC-LM3) were assessed by qRT-PCR and WB. ( G - H ) qRT-PCR and WB analysis demonstrating the mRNA and protein levels of Treg cell markers FOXP3 and CD25 in clinical samples. ( I ) ELISA quantification of TGF-β1 and IL-10 in HCC tumor and adjacent non-tumor tissues. ( J - K ) The levels of FOXP3 and CD25 in PBMCs co-cultured with FCN3-knockdown or FCN3-overexpressing HCC cells were evaluated by qRT-PCR and WB. ( L ) Flow cytometry analysis of CD4⁺CD25⁺FOXP3 + Treg cell proportion in PBMCs within the co-culture systems. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC/ OE-NC

Techniques Used: Expressing, Activation Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Knockdown, Flow Cytometry, Co-Culture Assay

FCN3 regulated Wnt/β-catenin signaling to influence Treg activation. ( A - B ) qRT-PCR and WB analysis of APC and β-catenin in HCC tumor and adjacent non-tumor tissues. ( C ) Clinical sample IF staining showing β-catenin expression and localization with DAPI nuclear counterstain. ( D - E ) The levels of APC and β-catenin in HCC cell lines were assessed by qRT-PCR and WB. ( F - G ) qRT-PCR and WB evaluation of APC and β-catenin in Hep3B cells treated with OE-NC, OE-FCN3, OE-FCN3 + DMSO, or OE-FCN3 + LY2090314 (β-catenin activator). ( H ) The levels of TGF-β1 and IL-10 in cell culture supernatant of each group were examined by ELISA. ( I - M ) Wound healing, Transwell and flow cytometry assays evaluating the effect of OE-FCN3 and OE-FCN3 + LY2090314 on cell migration, invasion and apoptosis of Hep3B cells. ( N ) Flow cytometry was performed to analyze the effects of OE-FCN3 and OE-FCN3 + LY2090314 treatment in Hep3B cells on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ OE-NC/ OE-FCN3 + DMSO
Figure Legend Snippet: FCN3 regulated Wnt/β-catenin signaling to influence Treg activation. ( A - B ) qRT-PCR and WB analysis of APC and β-catenin in HCC tumor and adjacent non-tumor tissues. ( C ) Clinical sample IF staining showing β-catenin expression and localization with DAPI nuclear counterstain. ( D - E ) The levels of APC and β-catenin in HCC cell lines were assessed by qRT-PCR and WB. ( F - G ) qRT-PCR and WB evaluation of APC and β-catenin in Hep3B cells treated with OE-NC, OE-FCN3, OE-FCN3 + DMSO, or OE-FCN3 + LY2090314 (β-catenin activator). ( H ) The levels of TGF-β1 and IL-10 in cell culture supernatant of each group were examined by ELISA. ( I - M ) Wound healing, Transwell and flow cytometry assays evaluating the effect of OE-FCN3 and OE-FCN3 + LY2090314 on cell migration, invasion and apoptosis of Hep3B cells. ( N ) Flow cytometry was performed to analyze the effects of OE-FCN3 and OE-FCN3 + LY2090314 treatment in Hep3B cells on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ OE-NC/ OE-FCN3 + DMSO

Techniques Used: Activation Assay, Quantitative RT-PCR, Staining, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Migration

Enhanced N-glycosylation of FCN3 in HCC. ( A ) Protein-protein interaction network obtained from BioGRID Database exhibited the connection between FCN3 and glycosylation-related proteins. ( B ) WB assay confirmed FCN3 band shift in HCC cell lysates, showing FCN3 galactosylated modification. ( C ) WB analysis of HCC cell lysates treated with PNGase F or O-Glycanase was performed to verify that FCN3 glycosylation was N-linked rather than O-linked. ( D ) FCN3 band shift in Hep3B cells treated with TM or DMSO was analyzed by WB. ( E ) WB analysis of FCN3 protein was performed in FCN3-knockdown HCC cells transfected with either Vector, FCN3-WT, or FCN3-N189Q. ( F - G ) IF staining and ELISA quantification showed the effects of the Asn189 mutation on FCN3 intracellular levels and its secretion into the culture supernatant, respectively
Figure Legend Snippet: Enhanced N-glycosylation of FCN3 in HCC. ( A ) Protein-protein interaction network obtained from BioGRID Database exhibited the connection between FCN3 and glycosylation-related proteins. ( B ) WB assay confirmed FCN3 band shift in HCC cell lysates, showing FCN3 galactosylated modification. ( C ) WB analysis of HCC cell lysates treated with PNGase F or O-Glycanase was performed to verify that FCN3 glycosylation was N-linked rather than O-linked. ( D ) FCN3 band shift in Hep3B cells treated with TM or DMSO was analyzed by WB. ( E ) WB analysis of FCN3 protein was performed in FCN3-knockdown HCC cells transfected with either Vector, FCN3-WT, or FCN3-N189Q. ( F - G ) IF staining and ELISA quantification showed the effects of the Asn189 mutation on FCN3 intracellular levels and its secretion into the culture supernatant, respectively

Techniques Used: Glycoproteomics, Electrophoretic Mobility Shift Assay, Modification, Knockdown, Transfection, Plasmid Preparation, Staining, Enzyme-linked Immunosorbent Assay, Mutagenesis

N-glycosylated FCN3 influenced Wnt/β-catenin signaling and Treg activation. ( A ) qRT-PCR analysis of APC and β-catenin mRNA levels in Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( B ) WB analysis showing the levels of APC, β-catenin, and p-β-catenin. ( C ) Subcellular localization of β-catenin was analyzed by WB. ( D ) ELISA was conducted to assess the levels of TGF-β1 and IL-10 in cell culture supernatant of Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( E - K ) The effects of FCN3 glycosylation on cell viability, migration, invasion and apoptosis in Hep3B cells were respectively evaluated by CCK-8, wound healing, Transwell and flow cytometry assays. ( L ) Flow cytometry demonstrating the impact of FCN3 glycosylation on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. FCN3-WT
Figure Legend Snippet: N-glycosylated FCN3 influenced Wnt/β-catenin signaling and Treg activation. ( A ) qRT-PCR analysis of APC and β-catenin mRNA levels in Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( B ) WB analysis showing the levels of APC, β-catenin, and p-β-catenin. ( C ) Subcellular localization of β-catenin was analyzed by WB. ( D ) ELISA was conducted to assess the levels of TGF-β1 and IL-10 in cell culture supernatant of Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( E - K ) The effects of FCN3 glycosylation on cell viability, migration, invasion and apoptosis in Hep3B cells were respectively evaluated by CCK-8, wound healing, Transwell and flow cytometry assays. ( L ) Flow cytometry demonstrating the impact of FCN3 glycosylation on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. FCN3-WT

Techniques Used: Activation Assay, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay, Cell Culture, Glycoproteomics, Migration, CCK-8 Assay, Flow Cytometry

STT3A involved in HCC progression via promoting FCN3 N-glycosylation. ( A ) Bioinformatics analysis revealing STT3A expression in normal and HCC liver tissues. ( B - D ) qRT-PCR, WB and IHC assays were performed to assess STT3A levels in HCC tumor and adjacent non-tumor tissues. ( E - F ) The mRNA and protein levels of STT3A in normal hepatocyte and HCC cell lines were analyzed by qRT-PCR and WB. ( G ) Correlation analysis between STT3A protein expression and FCN3 protein expression in tumor tissues from HCC patients. ( H ) Co-IP assay verified STT3A-FCN3 interaction. ( I ) WB analysis of FCN3 glycosylation in Hep3B cells transfected with sh-STT3A or sh-NC. ( J - K ) The effects of sh-STT3A on the expression of APC and β-catenin in Hep3B cells were examined by qRT-PCR and WB. ( L ) ELISA showing TGF-β1 and IL-10 levels in Hep3B cell supernatants following sh-STT3A or sh-NC treatment. ( M - Q ) Wound healing, Transwell and flow cytometry assays were conducted to evaluate the effects of sh-STT3A on Hep3B cell migration, invasion and apoptosis. ( R ) The influence of sh-STT3A on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures was assessed by flow cytometry. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC
Figure Legend Snippet: STT3A involved in HCC progression via promoting FCN3 N-glycosylation. ( A ) Bioinformatics analysis revealing STT3A expression in normal and HCC liver tissues. ( B - D ) qRT-PCR, WB and IHC assays were performed to assess STT3A levels in HCC tumor and adjacent non-tumor tissues. ( E - F ) The mRNA and protein levels of STT3A in normal hepatocyte and HCC cell lines were analyzed by qRT-PCR and WB. ( G ) Correlation analysis between STT3A protein expression and FCN3 protein expression in tumor tissues from HCC patients. ( H ) Co-IP assay verified STT3A-FCN3 interaction. ( I ) WB analysis of FCN3 glycosylation in Hep3B cells transfected with sh-STT3A or sh-NC. ( J - K ) The effects of sh-STT3A on the expression of APC and β-catenin in Hep3B cells were examined by qRT-PCR and WB. ( L ) ELISA showing TGF-β1 and IL-10 levels in Hep3B cell supernatants following sh-STT3A or sh-NC treatment. ( M - Q ) Wound healing, Transwell and flow cytometry assays were conducted to evaluate the effects of sh-STT3A on Hep3B cell migration, invasion and apoptosis. ( R ) The influence of sh-STT3A on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures was assessed by flow cytometry. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC

Techniques Used: Glycoproteomics, Expressing, Quantitative RT-PCR, Co-Immunoprecipitation Assay, Transfection, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Migration

STT3A promoted HCC development by regulating Treg cell activation in vivo. ( A ) Representative tumors of C57BL/6 mice at day 21 post-inoculation of Hepa1-6 cells with sh-STT3A or sh-NC transfection. ( B - C ) Tumor volume and body weight recorded during 21 days post Hepa1-6 cell inoculation demonstrated the effect of sh-STT3A on HCC tumor growth. ( D - F ) qRT-PCR and WB analysis of FOXP3 and CD25 in tumor tissues from HCC mice with sh-STT3A or sh-NC. ( G ) H&E-stained lung sections showing the impact of sh-STT3A on the number of metastatic nodules in HCC mice. ( H - J ) Tumor growth evaluation (images, volume, weight) in mice receiving Hepa1-6 cells with OE-NC, OE-STT3A, or OE-STT3A + DT. ( K - L ) ELISA assessing the influence of OE-STT3A and OE-STT3A + DT on TGF-β1 and IL-10 in HCC mouse tumor tissues. ( M ) Assessment of lung metastasis in HCC mice with OE-NC, OE-STT3A, or OE-STT3A + DT treatment using H&E staining. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sh-NC/ OE-NC/ OE-STT3A
Figure Legend Snippet: STT3A promoted HCC development by regulating Treg cell activation in vivo. ( A ) Representative tumors of C57BL/6 mice at day 21 post-inoculation of Hepa1-6 cells with sh-STT3A or sh-NC transfection. ( B - C ) Tumor volume and body weight recorded during 21 days post Hepa1-6 cell inoculation demonstrated the effect of sh-STT3A on HCC tumor growth. ( D - F ) qRT-PCR and WB analysis of FOXP3 and CD25 in tumor tissues from HCC mice with sh-STT3A or sh-NC. ( G ) H&E-stained lung sections showing the impact of sh-STT3A on the number of metastatic nodules in HCC mice. ( H - J ) Tumor growth evaluation (images, volume, weight) in mice receiving Hepa1-6 cells with OE-NC, OE-STT3A, or OE-STT3A + DT. ( K - L ) ELISA assessing the influence of OE-STT3A and OE-STT3A + DT on TGF-β1 and IL-10 in HCC mouse tumor tissues. ( M ) Assessment of lung metastasis in HCC mice with OE-NC, OE-STT3A, or OE-STT3A + DT treatment using H&E staining. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sh-NC/ OE-NC/ OE-STT3A

Techniques Used: Activation Assay, In Vivo, Transfection, Quantitative RT-PCR, Staining, Enzyme-linked Immunosorbent Assay

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Enzyme-linked Immunosorbent Assay:

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin
Article Snippet: Immunosuppressive cytokine levels were quantified using commercial ELISA kits (MultiSciences, Hangzhou, Zhejiang, China) including TGF-β1 ELISA kit (#EK981) and IL-10 ELISA kit (#EK210) according to the manufacturer’s protocol. .. For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions. ..



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Low FCN3 expression in HCC affected Treg cell activation. ( A ) Kaplan-Meier survival analysis showing the correlation between FCN3 levels and overall survival of HCC patients. ( B - D ) qRT-PCR, WB and IHC analysis of FCN3 expression in tumor ( T ) and adjacent non-tumor ( N ) tissues from HCC patients. ( E - F ) The levels of FCN3 in normal hepatocyte THLE-2 and HCC cell lines (HepG2, Hep3B, HCC-LM3) were assessed by qRT-PCR and WB. ( G - H ) qRT-PCR and WB analysis demonstrating the mRNA and protein levels of Treg cell markers FOXP3 and CD25 in clinical samples. ( I ) ELISA quantification of TGF-β1 and IL-10 in HCC tumor and adjacent non-tumor tissues. ( J - K ) The levels of FOXP3 and CD25 in PBMCs co-cultured with FCN3-knockdown or FCN3-overexpressing HCC cells were evaluated by qRT-PCR and WB. ( L ) Flow cytometry analysis of CD4⁺CD25⁺FOXP3 + Treg cell proportion in PBMCs within the co-culture systems. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC/ OE-NC

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: Low FCN3 expression in HCC affected Treg cell activation. ( A ) Kaplan-Meier survival analysis showing the correlation between FCN3 levels and overall survival of HCC patients. ( B - D ) qRT-PCR, WB and IHC analysis of FCN3 expression in tumor ( T ) and adjacent non-tumor ( N ) tissues from HCC patients. ( E - F ) The levels of FCN3 in normal hepatocyte THLE-2 and HCC cell lines (HepG2, Hep3B, HCC-LM3) were assessed by qRT-PCR and WB. ( G - H ) qRT-PCR and WB analysis demonstrating the mRNA and protein levels of Treg cell markers FOXP3 and CD25 in clinical samples. ( I ) ELISA quantification of TGF-β1 and IL-10 in HCC tumor and adjacent non-tumor tissues. ( J - K ) The levels of FOXP3 and CD25 in PBMCs co-cultured with FCN3-knockdown or FCN3-overexpressing HCC cells were evaluated by qRT-PCR and WB. ( L ) Flow cytometry analysis of CD4⁺CD25⁺FOXP3 + Treg cell proportion in PBMCs within the co-culture systems. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC/ OE-NC

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Knockdown, Flow Cytometry, Co-Culture Assay

FCN3 regulated Wnt/β-catenin signaling to influence Treg activation. ( A - B ) qRT-PCR and WB analysis of APC and β-catenin in HCC tumor and adjacent non-tumor tissues. ( C ) Clinical sample IF staining showing β-catenin expression and localization with DAPI nuclear counterstain. ( D - E ) The levels of APC and β-catenin in HCC cell lines were assessed by qRT-PCR and WB. ( F - G ) qRT-PCR and WB evaluation of APC and β-catenin in Hep3B cells treated with OE-NC, OE-FCN3, OE-FCN3 + DMSO, or OE-FCN3 + LY2090314 (β-catenin activator). ( H ) The levels of TGF-β1 and IL-10 in cell culture supernatant of each group were examined by ELISA. ( I - M ) Wound healing, Transwell and flow cytometry assays evaluating the effect of OE-FCN3 and OE-FCN3 + LY2090314 on cell migration, invasion and apoptosis of Hep3B cells. ( N ) Flow cytometry was performed to analyze the effects of OE-FCN3 and OE-FCN3 + LY2090314 treatment in Hep3B cells on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ OE-NC/ OE-FCN3 + DMSO

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: FCN3 regulated Wnt/β-catenin signaling to influence Treg activation. ( A - B ) qRT-PCR and WB analysis of APC and β-catenin in HCC tumor and adjacent non-tumor tissues. ( C ) Clinical sample IF staining showing β-catenin expression and localization with DAPI nuclear counterstain. ( D - E ) The levels of APC and β-catenin in HCC cell lines were assessed by qRT-PCR and WB. ( F - G ) qRT-PCR and WB evaluation of APC and β-catenin in Hep3B cells treated with OE-NC, OE-FCN3, OE-FCN3 + DMSO, or OE-FCN3 + LY2090314 (β-catenin activator). ( H ) The levels of TGF-β1 and IL-10 in cell culture supernatant of each group were examined by ELISA. ( I - M ) Wound healing, Transwell and flow cytometry assays evaluating the effect of OE-FCN3 and OE-FCN3 + LY2090314 on cell migration, invasion and apoptosis of Hep3B cells. ( N ) Flow cytometry was performed to analyze the effects of OE-FCN3 and OE-FCN3 + LY2090314 treatment in Hep3B cells on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ OE-NC/ OE-FCN3 + DMSO

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Activation Assay, Quantitative RT-PCR, Staining, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Migration

Enhanced N-glycosylation of FCN3 in HCC. ( A ) Protein-protein interaction network obtained from BioGRID Database exhibited the connection between FCN3 and glycosylation-related proteins. ( B ) WB assay confirmed FCN3 band shift in HCC cell lysates, showing FCN3 galactosylated modification. ( C ) WB analysis of HCC cell lysates treated with PNGase F or O-Glycanase was performed to verify that FCN3 glycosylation was N-linked rather than O-linked. ( D ) FCN3 band shift in Hep3B cells treated with TM or DMSO was analyzed by WB. ( E ) WB analysis of FCN3 protein was performed in FCN3-knockdown HCC cells transfected with either Vector, FCN3-WT, or FCN3-N189Q. ( F - G ) IF staining and ELISA quantification showed the effects of the Asn189 mutation on FCN3 intracellular levels and its secretion into the culture supernatant, respectively

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: Enhanced N-glycosylation of FCN3 in HCC. ( A ) Protein-protein interaction network obtained from BioGRID Database exhibited the connection between FCN3 and glycosylation-related proteins. ( B ) WB assay confirmed FCN3 band shift in HCC cell lysates, showing FCN3 galactosylated modification. ( C ) WB analysis of HCC cell lysates treated with PNGase F or O-Glycanase was performed to verify that FCN3 glycosylation was N-linked rather than O-linked. ( D ) FCN3 band shift in Hep3B cells treated with TM or DMSO was analyzed by WB. ( E ) WB analysis of FCN3 protein was performed in FCN3-knockdown HCC cells transfected with either Vector, FCN3-WT, or FCN3-N189Q. ( F - G ) IF staining and ELISA quantification showed the effects of the Asn189 mutation on FCN3 intracellular levels and its secretion into the culture supernatant, respectively

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Glycoproteomics, Electrophoretic Mobility Shift Assay, Modification, Knockdown, Transfection, Plasmid Preparation, Staining, Enzyme-linked Immunosorbent Assay, Mutagenesis

N-glycosylated FCN3 influenced Wnt/β-catenin signaling and Treg activation. ( A ) qRT-PCR analysis of APC and β-catenin mRNA levels in Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( B ) WB analysis showing the levels of APC, β-catenin, and p-β-catenin. ( C ) Subcellular localization of β-catenin was analyzed by WB. ( D ) ELISA was conducted to assess the levels of TGF-β1 and IL-10 in cell culture supernatant of Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( E - K ) The effects of FCN3 glycosylation on cell viability, migration, invasion and apoptosis in Hep3B cells were respectively evaluated by CCK-8, wound healing, Transwell and flow cytometry assays. ( L ) Flow cytometry demonstrating the impact of FCN3 glycosylation on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. FCN3-WT

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: N-glycosylated FCN3 influenced Wnt/β-catenin signaling and Treg activation. ( A ) qRT-PCR analysis of APC and β-catenin mRNA levels in Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( B ) WB analysis showing the levels of APC, β-catenin, and p-β-catenin. ( C ) Subcellular localization of β-catenin was analyzed by WB. ( D ) ELISA was conducted to assess the levels of TGF-β1 and IL-10 in cell culture supernatant of Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( E - K ) The effects of FCN3 glycosylation on cell viability, migration, invasion and apoptosis in Hep3B cells were respectively evaluated by CCK-8, wound healing, Transwell and flow cytometry assays. ( L ) Flow cytometry demonstrating the impact of FCN3 glycosylation on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. FCN3-WT

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Activation Assay, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay, Cell Culture, Glycoproteomics, Migration, CCK-8 Assay, Flow Cytometry

STT3A involved in HCC progression via promoting FCN3 N-glycosylation. ( A ) Bioinformatics analysis revealing STT3A expression in normal and HCC liver tissues. ( B - D ) qRT-PCR, WB and IHC assays were performed to assess STT3A levels in HCC tumor and adjacent non-tumor tissues. ( E - F ) The mRNA and protein levels of STT3A in normal hepatocyte and HCC cell lines were analyzed by qRT-PCR and WB. ( G ) Correlation analysis between STT3A protein expression and FCN3 protein expression in tumor tissues from HCC patients. ( H ) Co-IP assay verified STT3A-FCN3 interaction. ( I ) WB analysis of FCN3 glycosylation in Hep3B cells transfected with sh-STT3A or sh-NC. ( J - K ) The effects of sh-STT3A on the expression of APC and β-catenin in Hep3B cells were examined by qRT-PCR and WB. ( L ) ELISA showing TGF-β1 and IL-10 levels in Hep3B cell supernatants following sh-STT3A or sh-NC treatment. ( M - Q ) Wound healing, Transwell and flow cytometry assays were conducted to evaluate the effects of sh-STT3A on Hep3B cell migration, invasion and apoptosis. ( R ) The influence of sh-STT3A on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures was assessed by flow cytometry. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: STT3A involved in HCC progression via promoting FCN3 N-glycosylation. ( A ) Bioinformatics analysis revealing STT3A expression in normal and HCC liver tissues. ( B - D ) qRT-PCR, WB and IHC assays were performed to assess STT3A levels in HCC tumor and adjacent non-tumor tissues. ( E - F ) The mRNA and protein levels of STT3A in normal hepatocyte and HCC cell lines were analyzed by qRT-PCR and WB. ( G ) Correlation analysis between STT3A protein expression and FCN3 protein expression in tumor tissues from HCC patients. ( H ) Co-IP assay verified STT3A-FCN3 interaction. ( I ) WB analysis of FCN3 glycosylation in Hep3B cells transfected with sh-STT3A or sh-NC. ( J - K ) The effects of sh-STT3A on the expression of APC and β-catenin in Hep3B cells were examined by qRT-PCR and WB. ( L ) ELISA showing TGF-β1 and IL-10 levels in Hep3B cell supernatants following sh-STT3A or sh-NC treatment. ( M - Q ) Wound healing, Transwell and flow cytometry assays were conducted to evaluate the effects of sh-STT3A on Hep3B cell migration, invasion and apoptosis. ( R ) The influence of sh-STT3A on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures was assessed by flow cytometry. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Glycoproteomics, Expressing, Quantitative RT-PCR, Co-Immunoprecipitation Assay, Transfection, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Migration

STT3A promoted HCC development by regulating Treg cell activation in vivo. ( A ) Representative tumors of C57BL/6 mice at day 21 post-inoculation of Hepa1-6 cells with sh-STT3A or sh-NC transfection. ( B - C ) Tumor volume and body weight recorded during 21 days post Hepa1-6 cell inoculation demonstrated the effect of sh-STT3A on HCC tumor growth. ( D - F ) qRT-PCR and WB analysis of FOXP3 and CD25 in tumor tissues from HCC mice with sh-STT3A or sh-NC. ( G ) H&E-stained lung sections showing the impact of sh-STT3A on the number of metastatic nodules in HCC mice. ( H - J ) Tumor growth evaluation (images, volume, weight) in mice receiving Hepa1-6 cells with OE-NC, OE-STT3A, or OE-STT3A + DT. ( K - L ) ELISA assessing the influence of OE-STT3A and OE-STT3A + DT on TGF-β1 and IL-10 in HCC mouse tumor tissues. ( M ) Assessment of lung metastasis in HCC mice with OE-NC, OE-STT3A, or OE-STT3A + DT treatment using H&E staining. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sh-NC/ OE-NC/ OE-STT3A

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: STT3A promoted HCC development by regulating Treg cell activation in vivo. ( A ) Representative tumors of C57BL/6 mice at day 21 post-inoculation of Hepa1-6 cells with sh-STT3A or sh-NC transfection. ( B - C ) Tumor volume and body weight recorded during 21 days post Hepa1-6 cell inoculation demonstrated the effect of sh-STT3A on HCC tumor growth. ( D - F ) qRT-PCR and WB analysis of FOXP3 and CD25 in tumor tissues from HCC mice with sh-STT3A or sh-NC. ( G ) H&E-stained lung sections showing the impact of sh-STT3A on the number of metastatic nodules in HCC mice. ( H - J ) Tumor growth evaluation (images, volume, weight) in mice receiving Hepa1-6 cells with OE-NC, OE-STT3A, or OE-STT3A + DT. ( K - L ) ELISA assessing the influence of OE-STT3A and OE-STT3A + DT on TGF-β1 and IL-10 in HCC mouse tumor tissues. ( M ) Assessment of lung metastasis in HCC mice with OE-NC, OE-STT3A, or OE-STT3A + DT treatment using H&E staining. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sh-NC/ OE-NC/ OE-STT3A

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Activation Assay, In Vivo, Transfection, Quantitative RT-PCR, Staining, Enzyme-linked Immunosorbent Assay

Low FCN3 expression in HCC affected Treg cell activation. ( A ) Kaplan-Meier survival analysis showing the correlation between FCN3 levels and overall survival of HCC patients. ( B - D ) qRT-PCR, WB and IHC analysis of FCN3 expression in tumor ( T ) and adjacent non-tumor ( N ) tissues from HCC patients. ( E - F ) The levels of FCN3 in normal hepatocyte THLE-2 and HCC cell lines (HepG2, Hep3B, HCC-LM3) were assessed by qRT-PCR and WB. ( G - H ) qRT-PCR and WB analysis demonstrating the mRNA and protein levels of Treg cell markers FOXP3 and CD25 in clinical samples. ( I ) ELISA quantification of TGF-β1 and IL-10 in HCC tumor and adjacent non-tumor tissues. ( J - K ) The levels of FOXP3 and CD25 in PBMCs co-cultured with FCN3-knockdown or FCN3-overexpressing HCC cells were evaluated by qRT-PCR and WB. ( L ) Flow cytometry analysis of CD4⁺CD25⁺FOXP3 + Treg cell proportion in PBMCs within the co-culture systems. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC/ OE-NC

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: Low FCN3 expression in HCC affected Treg cell activation. ( A ) Kaplan-Meier survival analysis showing the correlation between FCN3 levels and overall survival of HCC patients. ( B - D ) qRT-PCR, WB and IHC analysis of FCN3 expression in tumor ( T ) and adjacent non-tumor ( N ) tissues from HCC patients. ( E - F ) The levels of FCN3 in normal hepatocyte THLE-2 and HCC cell lines (HepG2, Hep3B, HCC-LM3) were assessed by qRT-PCR and WB. ( G - H ) qRT-PCR and WB analysis demonstrating the mRNA and protein levels of Treg cell markers FOXP3 and CD25 in clinical samples. ( I ) ELISA quantification of TGF-β1 and IL-10 in HCC tumor and adjacent non-tumor tissues. ( J - K ) The levels of FOXP3 and CD25 in PBMCs co-cultured with FCN3-knockdown or FCN3-overexpressing HCC cells were evaluated by qRT-PCR and WB. ( L ) Flow cytometry analysis of CD4⁺CD25⁺FOXP3 + Treg cell proportion in PBMCs within the co-culture systems. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC/ OE-NC

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Cell Culture, Knockdown, Flow Cytometry, Co-Culture Assay

FCN3 regulated Wnt/β-catenin signaling to influence Treg activation. ( A - B ) qRT-PCR and WB analysis of APC and β-catenin in HCC tumor and adjacent non-tumor tissues. ( C ) Clinical sample IF staining showing β-catenin expression and localization with DAPI nuclear counterstain. ( D - E ) The levels of APC and β-catenin in HCC cell lines were assessed by qRT-PCR and WB. ( F - G ) qRT-PCR and WB evaluation of APC and β-catenin in Hep3B cells treated with OE-NC, OE-FCN3, OE-FCN3 + DMSO, or OE-FCN3 + LY2090314 (β-catenin activator). ( H ) The levels of TGF-β1 and IL-10 in cell culture supernatant of each group were examined by ELISA. ( I - M ) Wound healing, Transwell and flow cytometry assays evaluating the effect of OE-FCN3 and OE-FCN3 + LY2090314 on cell migration, invasion and apoptosis of Hep3B cells. ( N ) Flow cytometry was performed to analyze the effects of OE-FCN3 and OE-FCN3 + LY2090314 treatment in Hep3B cells on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ OE-NC/ OE-FCN3 + DMSO

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: FCN3 regulated Wnt/β-catenin signaling to influence Treg activation. ( A - B ) qRT-PCR and WB analysis of APC and β-catenin in HCC tumor and adjacent non-tumor tissues. ( C ) Clinical sample IF staining showing β-catenin expression and localization with DAPI nuclear counterstain. ( D - E ) The levels of APC and β-catenin in HCC cell lines were assessed by qRT-PCR and WB. ( F - G ) qRT-PCR and WB evaluation of APC and β-catenin in Hep3B cells treated with OE-NC, OE-FCN3, OE-FCN3 + DMSO, or OE-FCN3 + LY2090314 (β-catenin activator). ( H ) The levels of TGF-β1 and IL-10 in cell culture supernatant of each group were examined by ELISA. ( I - M ) Wound healing, Transwell and flow cytometry assays evaluating the effect of OE-FCN3 and OE-FCN3 + LY2090314 on cell migration, invasion and apoptosis of Hep3B cells. ( N ) Flow cytometry was performed to analyze the effects of OE-FCN3 and OE-FCN3 + LY2090314 treatment in Hep3B cells on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ OE-NC/ OE-FCN3 + DMSO

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Activation Assay, Quantitative RT-PCR, Staining, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Migration

Enhanced N-glycosylation of FCN3 in HCC. ( A ) Protein-protein interaction network obtained from BioGRID Database exhibited the connection between FCN3 and glycosylation-related proteins. ( B ) WB assay confirmed FCN3 band shift in HCC cell lysates, showing FCN3 galactosylated modification. ( C ) WB analysis of HCC cell lysates treated with PNGase F or O-Glycanase was performed to verify that FCN3 glycosylation was N-linked rather than O-linked. ( D ) FCN3 band shift in Hep3B cells treated with TM or DMSO was analyzed by WB. ( E ) WB analysis of FCN3 protein was performed in FCN3-knockdown HCC cells transfected with either Vector, FCN3-WT, or FCN3-N189Q. ( F - G ) IF staining and ELISA quantification showed the effects of the Asn189 mutation on FCN3 intracellular levels and its secretion into the culture supernatant, respectively

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: Enhanced N-glycosylation of FCN3 in HCC. ( A ) Protein-protein interaction network obtained from BioGRID Database exhibited the connection between FCN3 and glycosylation-related proteins. ( B ) WB assay confirmed FCN3 band shift in HCC cell lysates, showing FCN3 galactosylated modification. ( C ) WB analysis of HCC cell lysates treated with PNGase F or O-Glycanase was performed to verify that FCN3 glycosylation was N-linked rather than O-linked. ( D ) FCN3 band shift in Hep3B cells treated with TM or DMSO was analyzed by WB. ( E ) WB analysis of FCN3 protein was performed in FCN3-knockdown HCC cells transfected with either Vector, FCN3-WT, or FCN3-N189Q. ( F - G ) IF staining and ELISA quantification showed the effects of the Asn189 mutation on FCN3 intracellular levels and its secretion into the culture supernatant, respectively

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Glycoproteomics, Electrophoretic Mobility Shift Assay, Modification, Knockdown, Transfection, Plasmid Preparation, Staining, Enzyme-linked Immunosorbent Assay, Mutagenesis

N-glycosylated FCN3 influenced Wnt/β-catenin signaling and Treg activation. ( A ) qRT-PCR analysis of APC and β-catenin mRNA levels in Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( B ) WB analysis showing the levels of APC, β-catenin, and p-β-catenin. ( C ) Subcellular localization of β-catenin was analyzed by WB. ( D ) ELISA was conducted to assess the levels of TGF-β1 and IL-10 in cell culture supernatant of Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( E - K ) The effects of FCN3 glycosylation on cell viability, migration, invasion and apoptosis in Hep3B cells were respectively evaluated by CCK-8, wound healing, Transwell and flow cytometry assays. ( L ) Flow cytometry demonstrating the impact of FCN3 glycosylation on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. FCN3-WT

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: N-glycosylated FCN3 influenced Wnt/β-catenin signaling and Treg activation. ( A ) qRT-PCR analysis of APC and β-catenin mRNA levels in Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( B ) WB analysis showing the levels of APC, β-catenin, and p-β-catenin. ( C ) Subcellular localization of β-catenin was analyzed by WB. ( D ) ELISA was conducted to assess the levels of TGF-β1 and IL-10 in cell culture supernatant of Hep3B cells transfected with FCN3-WT or FCN3-N189Q. ( E - K ) The effects of FCN3 glycosylation on cell viability, migration, invasion and apoptosis in Hep3B cells were respectively evaluated by CCK-8, wound healing, Transwell and flow cytometry assays. ( L ) Flow cytometry demonstrating the impact of FCN3 glycosylation on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. FCN3-WT

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Activation Assay, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay, Cell Culture, Glycoproteomics, Migration, CCK-8 Assay, Flow Cytometry

STT3A involved in HCC progression via promoting FCN3 N-glycosylation. ( A ) Bioinformatics analysis revealing STT3A expression in normal and HCC liver tissues. ( B - D ) qRT-PCR, WB and IHC assays were performed to assess STT3A levels in HCC tumor and adjacent non-tumor tissues. ( E - F ) The mRNA and protein levels of STT3A in normal hepatocyte and HCC cell lines were analyzed by qRT-PCR and WB. ( G ) Correlation analysis between STT3A protein expression and FCN3 protein expression in tumor tissues from HCC patients. ( H ) Co-IP assay verified STT3A-FCN3 interaction. ( I ) WB analysis of FCN3 glycosylation in Hep3B cells transfected with sh-STT3A or sh-NC. ( J - K ) The effects of sh-STT3A on the expression of APC and β-catenin in Hep3B cells were examined by qRT-PCR and WB. ( L ) ELISA showing TGF-β1 and IL-10 levels in Hep3B cell supernatants following sh-STT3A or sh-NC treatment. ( M - Q ) Wound healing, Transwell and flow cytometry assays were conducted to evaluate the effects of sh-STT3A on Hep3B cell migration, invasion and apoptosis. ( R ) The influence of sh-STT3A on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures was assessed by flow cytometry. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: STT3A involved in HCC progression via promoting FCN3 N-glycosylation. ( A ) Bioinformatics analysis revealing STT3A expression in normal and HCC liver tissues. ( B - D ) qRT-PCR, WB and IHC assays were performed to assess STT3A levels in HCC tumor and adjacent non-tumor tissues. ( E - F ) The mRNA and protein levels of STT3A in normal hepatocyte and HCC cell lines were analyzed by qRT-PCR and WB. ( G ) Correlation analysis between STT3A protein expression and FCN3 protein expression in tumor tissues from HCC patients. ( H ) Co-IP assay verified STT3A-FCN3 interaction. ( I ) WB analysis of FCN3 glycosylation in Hep3B cells transfected with sh-STT3A or sh-NC. ( J - K ) The effects of sh-STT3A on the expression of APC and β-catenin in Hep3B cells were examined by qRT-PCR and WB. ( L ) ELISA showing TGF-β1 and IL-10 levels in Hep3B cell supernatants following sh-STT3A or sh-NC treatment. ( M - Q ) Wound healing, Transwell and flow cytometry assays were conducted to evaluate the effects of sh-STT3A on Hep3B cell migration, invasion and apoptosis. ( R ) The influence of sh-STT3A on the proportion of CD4⁺CD25⁺FOXP3 + Treg cells in Hep3B-PBMC co-cultures was assessed by flow cytometry. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. N/ THLE-2/ sh-NC

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Glycoproteomics, Expressing, Quantitative RT-PCR, Co-Immunoprecipitation Assay, Transfection, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Migration

STT3A promoted HCC development by regulating Treg cell activation in vivo. ( A ) Representative tumors of C57BL/6 mice at day 21 post-inoculation of Hepa1-6 cells with sh-STT3A or sh-NC transfection. ( B - C ) Tumor volume and body weight recorded during 21 days post Hepa1-6 cell inoculation demonstrated the effect of sh-STT3A on HCC tumor growth. ( D - F ) qRT-PCR and WB analysis of FOXP3 and CD25 in tumor tissues from HCC mice with sh-STT3A or sh-NC. ( G ) H&E-stained lung sections showing the impact of sh-STT3A on the number of metastatic nodules in HCC mice. ( H - J ) Tumor growth evaluation (images, volume, weight) in mice receiving Hepa1-6 cells with OE-NC, OE-STT3A, or OE-STT3A + DT. ( K - L ) ELISA assessing the influence of OE-STT3A and OE-STT3A + DT on TGF-β1 and IL-10 in HCC mouse tumor tissues. ( M ) Assessment of lung metastasis in HCC mice with OE-NC, OE-STT3A, or OE-STT3A + DT treatment using H&E staining. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sh-NC/ OE-NC/ OE-STT3A

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: STT3A-mediated FCN3 N-glycosylation promotes Treg cell activation to drive hepatocellular carcinoma progression via Wnt/β-catenin

doi: 10.1007/s13402-025-01159-1

Figure Lengend Snippet: STT3A promoted HCC development by regulating Treg cell activation in vivo. ( A ) Representative tumors of C57BL/6 mice at day 21 post-inoculation of Hepa1-6 cells with sh-STT3A or sh-NC transfection. ( B - C ) Tumor volume and body weight recorded during 21 days post Hepa1-6 cell inoculation demonstrated the effect of sh-STT3A on HCC tumor growth. ( D - F ) qRT-PCR and WB analysis of FOXP3 and CD25 in tumor tissues from HCC mice with sh-STT3A or sh-NC. ( G ) H&E-stained lung sections showing the impact of sh-STT3A on the number of metastatic nodules in HCC mice. ( H - J ) Tumor growth evaluation (images, volume, weight) in mice receiving Hepa1-6 cells with OE-NC, OE-STT3A, or OE-STT3A + DT. ( K - L ) ELISA assessing the influence of OE-STT3A and OE-STT3A + DT on TGF-β1 and IL-10 in HCC mouse tumor tissues. ( M ) Assessment of lung metastasis in HCC mice with OE-NC, OE-STT3A, or OE-STT3A + DT treatment using H&E staining. *** p < 0.001, ** p < 0.01, * p < 0.05 vs. sh-NC/ OE-NC/ OE-STT3A

Article Snippet: For detection of secreted FCN3, the Human FCN3 ELISA Kit (#CSB‐E13104h, Cusabio, Wuhan, Hubei, China) was used following the supplier’s instructions.

Techniques: Activation Assay, In Vivo, Transfection, Quantitative RT-PCR, Staining, Enzyme-linked Immunosorbent Assay

The summed normalized weights of 20 differently expressed genes (DEGs) in six classifications machine learnings (MLs).

Journal: Frontiers in Cardiovascular Medicine

Article Title: Multiple-model machine learning identifies potential functional genes in dilated cardiomyopathy

doi: 10.3389/fcvm.2022.1044443

Figure Lengend Snippet: The summed normalized weights of 20 differently expressed genes (DEGs) in six classifications machine learnings (MLs).

Article Snippet: All the plasma levels of SERPINA3, FCN3, and FRZB were measured by ELISA kits (SERPINA3 Human ELISA Kit, Abcam, Cambridge, UK; Hycult Biotechnology, Uden, The Netherlands; R&D Systems, Minneapolis, MN, USA, respectively).

Techniques:

The receiver operating characteristic (ROC) of SERPINA3, FRZB, and FCN3 between the control and dilated cardiomyopathy (DCM) groups. (A,C,E) The ROC of SERPINA3, FRZB, and FCN3 in the Training set; (B,D,F) the ROC of SERPINA3, FRZB, and FCN3 in the testing set.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Multiple-model machine learning identifies potential functional genes in dilated cardiomyopathy

doi: 10.3389/fcvm.2022.1044443

Figure Lengend Snippet: The receiver operating characteristic (ROC) of SERPINA3, FRZB, and FCN3 between the control and dilated cardiomyopathy (DCM) groups. (A,C,E) The ROC of SERPINA3, FRZB, and FCN3 in the Training set; (B,D,F) the ROC of SERPINA3, FRZB, and FCN3 in the testing set.

Article Snippet: All the plasma levels of SERPINA3, FCN3, and FRZB were measured by ELISA kits (SERPINA3 Human ELISA Kit, Abcam, Cambridge, UK; Hycult Biotechnology, Uden, The Netherlands; R&D Systems, Minneapolis, MN, USA, respectively).

Techniques: Control

The immune correlation between diagnosis genes and immune cells. (A–C) The lollipop map and linear regression map in SERPINA3, FRAB, and FCN3.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Multiple-model machine learning identifies potential functional genes in dilated cardiomyopathy

doi: 10.3389/fcvm.2022.1044443

Figure Lengend Snippet: The immune correlation between diagnosis genes and immune cells. (A–C) The lollipop map and linear regression map in SERPINA3, FRAB, and FCN3.

Article Snippet: All the plasma levels of SERPINA3, FCN3, and FRZB were measured by ELISA kits (SERPINA3 Human ELISA Kit, Abcam, Cambridge, UK; Hycult Biotechnology, Uden, The Netherlands; R&D Systems, Minneapolis, MN, USA, respectively).

Techniques:

The plasma levels of SERPINA3, FRZB, and FCN3 in dilated cardiomyopathy (DCM) patients and healthy controls. ***Mean P < 0.001.

Journal: Frontiers in Cardiovascular Medicine

Article Title: Multiple-model machine learning identifies potential functional genes in dilated cardiomyopathy

doi: 10.3389/fcvm.2022.1044443

Figure Lengend Snippet: The plasma levels of SERPINA3, FRZB, and FCN3 in dilated cardiomyopathy (DCM) patients and healthy controls. ***Mean P < 0.001.

Article Snippet: All the plasma levels of SERPINA3, FCN3, and FRZB were measured by ELISA kits (SERPINA3 Human ELISA Kit, Abcam, Cambridge, UK; Hycult Biotechnology, Uden, The Netherlands; R&D Systems, Minneapolis, MN, USA, respectively).

Techniques: