Journal: Cell Death & Disease

Article Title: HHIPL2 positively governs Hedgehog signaling to accelerate non-small cell lung cancer progression via enhancing HNRNPC-mediated HNF1A mRNA stabilization

doi: 10.1038/s41419-025-08331-3

Figure Lengend Snippet: A GLI luciferase assays of A549, H157, and H460 cells with HHIP, HHIPL1, and HHIPL2 overexpression. The GLI luciferase activity is normalized to Renilla ( n = 3 in each group). B GLI luciferase assays of A549, H157, and H460 cells with HHIPL2 knockdown. The GLI luciferase activity is normalized to Renilla ( n = 3 in each group). Relative RT-qPCR analysis of the mRNA levels of the genes related to the Hedgehog signaling in A549 ( C ), H157 ( D ), and H460 ( E ) cells with or without HHIPL2 knockdown ( n = 3 in each group). F Knockdown of HHIPL2 in A549, H157, and H460 cells. Cell lysates were analyzed by Western blot. G GLI luciferase assays of A549, H157, and H460 cells overexpressing HHIPL2 and SHH knockdown. The GLI luciferase activity is normalized to Renilla ( n = 3 in each group). Relative RT-qPCR analysis of the mRNA levels of the genes related to the Hedgehog signaling in A549 ( H ), H157 ( I ), and H460 ( J ) cells overexpressing HHIPL2 and SHH knockdown ( n = 3 in each group). K Overexpression of HHIPL2 in A549, H157, and H460 cells with or without SHH knockdown. Cell lysates were analyzed by Western blot. L Schematic showing that HHIPL2, possibly via an unknown mechanism, regulates Sonic Hedgehog signaling. Sonic Hedgehog signaling is roughly as follows: in the absence of SHH, PTCH1 suppresses the activity of SMO, which is released upon SHH binding to PTCH1, allowing SMO to activate its downstream targets. GLI, as the transcription factor, translocates into the nucleus to promote the transcription of related genes, thereby activating Sonic Hedgehog signaling. Data in ( A – E, G – J ) are presented as the mean ± SD. Statistical significance was assessed by a one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001. Experiments ( A – K ) were repeated at least three times.

Article Snippet: Intriguingly, data from The Human Protein Atlas online database showed that mRNA expression of HHIPL2 was upregulated in the majority of human cancer cell lines compared to non-cancerous cell lines (Fig. ), indicating that higher expression of HHIPL2 is widespread in a pan-cancer manner.

Techniques: Luciferase, Over Expression, Activity Assay, Knockdown, Quantitative RT-PCR, Western Blot, Binding Assay

Journal: Cell Death & Disease

Article Title: HHIPL2 positively governs Hedgehog signaling to accelerate non-small cell lung cancer progression via enhancing HNRNPC-mediated HNF1A mRNA stabilization

doi: 10.1038/s41419-025-08331-3

Figure Lengend Snippet: A Representative HHIPL2 IHC images of NSCLC tumor tissues and normal tissues ( n = 55 per group). Scale bars as shown. B HHIPL2 IHC scores in NSCLC tumor tissues and normal tissues. HHIPL2 IHC scores of NSCLC tissues were compared between T1-2 and T3-4 ( C ) or Stage I and II-III ( D ). Kaplan–Meier plots of the overall survival of Lung adenocarcinoma patients ( E ), Lung squamous cell carcinoma patients ( F ), and Lung carcinoma patients ( G ) stratified by HHIPL2 expression. The data were acquired from the Kaplan-Meier plotter database. H Knockdown of HHIPL2 in A549 and H460 cells. Cell lysates were analyzed by western blotting with the indicated antibodies. I Colony formation assays in A549 and H460 cells with or without HHIPL2 knockdown. ImageJ was used to perform quantitative analysis ( n = 3 in each group). J , K CCK-8 assays in A549 and H460 cells with or without HHIPL2 knockdown ( n = 3 in each group). L , M Effects of HHIPL2 knockdown on migration and invasion in A549 and H460 cells using transwell assays. ImageJ was used to perform quantitative analysis ( n = 4 in each group). Scale bar, 100 μm. Representative images of xenograft tumors after subcutaneous injection of A549 cells with HHIPL2 knockdown and controls ( N ), Tumor weights ( O ) and tumor volumes ( P ) were measured ( n = 5 per group). Representative images of lung metastasis models in nude mice after tail injection of A549 cells with HHIPL2 knockdown ( Q ) and quantification of pulmonary metastatic nodules ( R ) ( n = 5 per group). Data in ( B – D , I – M , O , P , R ) are presented as the mean ± SD. Statistical significance was assessed by the Wilcoxon matched-pairs signed rank test ( B ), the Mann–Whitney test ( C , D ), a one-way ANOVA ( I , L , M , O , R ), and a two-way ANOVA ( J , K , P ). ** P < 0.01, *** P < 0.001. Experiments ( H – M ) were repeated at least three times.

Article Snippet: Intriguingly, data from The Human Protein Atlas online database showed that mRNA expression of HHIPL2 was upregulated in the majority of human cancer cell lines compared to non-cancerous cell lines (Fig. ), indicating that higher expression of HHIPL2 is widespread in a pan-cancer manner.

Techniques: Expressing, Knockdown, Western Blot, CCK-8 Assay, Migration, Injection, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: HHIPL2 positively governs Hedgehog signaling to accelerate non-small cell lung cancer progression via enhancing HNRNPC-mediated HNF1A mRNA stabilization

doi: 10.1038/s41419-025-08331-3

Figure Lengend Snippet: A Overexpression of HHIPL2 in A549 and H157 cells with or without SHH knockdown. Cell lysates were analyzed by Western blot. B Effects of HHIPL2 overexpression with or without SHH knockdown in A549 and H157 cells using colony formation assays. ImageJ was used to perform quantitative analysis ( n = 3 in each group). Effects of HHIPL2 overexpression with or without SHH knockdown in A549 ( C ) and H157 ( D ) cells using CCK-8 assays ( n = 3 in each group). Effects of HHIPL2 overexpression with or without SHH knockdown on migration and invasion in A549 ( E ) and H157 ( F ) cells using transwell assays. ImageJ was used to perform quantitative analysis ( n = 4 in each group). Scale bar, 100 μm. G HHIPL2 expression was highly correlated with SHH expression in NSCLC tissues analyzed by IHC staining. Scale bars as shown. H Spearman correlation plot of HHIPL2 and SHH IHC scores in 55 NSCLC tissue samples. I Schematic showing that HHIPL2 accelerates NSCLC progression by regulating Sonic Hedgehog signaling. Data in ( B – F ) are presented as the mean ± SD. Statistical significance was assessed by a one-way ANOVA( B , E , F ), a two-way ANOVA ( C , D ), and a Spearman’s rank correlation coefficient analysis ( H ). * P < 0.05, ** P < 0.01, *** P < 0.001. Experiments ( A – F ) were repeated at least three times.

Article Snippet: Intriguingly, data from The Human Protein Atlas online database showed that mRNA expression of HHIPL2 was upregulated in the majority of human cancer cell lines compared to non-cancerous cell lines (Fig. ), indicating that higher expression of HHIPL2 is widespread in a pan-cancer manner.

Techniques: Over Expression, Knockdown, Western Blot, CCK-8 Assay, Migration, Expressing, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: HHIPL2 positively governs Hedgehog signaling to accelerate non-small cell lung cancer progression via enhancing HNRNPC-mediated HNF1A mRNA stabilization

doi: 10.1038/s41419-025-08331-3

Figure Lengend Snippet: A The dual-luciferase reporter assays of A549 and H460 cells analysis of the SHH promoter activity with HHIPL2 knockdown. The luciferase activity is normalized to Renilla ( n = 3 in each group). B The dual-luciferase reporter assays of A549 and H157 cells analysis of the SHH promoter activity with HHIPL2 overexpression. The luciferase activity is normalized to Renilla ( n = 3 in each group). C Schematic showing that the screening process of three transcription factors (TFs) binds the SHH promoter region. D Relative RT-qPCR analysis of the mRNA levels of HNF1A in A549 and H460 cells with or without HHIPL2 knockdown ( n = 3 in each group). Relative RT-qPCR analysis of the mRNA levels of the genes related to the Sonic Hedgehog pathway in A549 ( E ) and H157 cells ( F ) with HHIPL2 overexpression and HNF1A knockdown ( n = 3 in each group). G Overexpression of HHIPL2 in A549 and H157 cells with or without HNF1A knockdown. Cell lysates were analyzed by Western blot. H GLI luciferase assays of A549 and H157 cells with HHIPL2 overexpression and HNF1A knockdown. The GLI luciferase activity is normalized to Renilla ( n = 3 in each group). I Effects of HHIPL2 overexpression with or without HNF1A knockdown in A549 and H157 cells using colony formation assays. ImageJ was used to perform quantitative analysis ( n = 3 in each group). J Effects of HHIPL2 overexpression with or without HNF1A knockdown in A549 cells using CCK-8 assays ( n = 3 in each group). K Effects of HHIPL2 overexpression with or without HNF1A knockdown on migration and invasion in A549 cells using transwell assays. ImageJ was used to perform quantitative analysis ( n = 4 in each group). Scale bar, 100 μm. L HHIPL2 expression was highly correlated with HNF1A expression in NSCLC tissues analyzed by IHC staining. Scale bars as shown. M Spearman correlation plot of HHIPL2 and HNF1A IHC scores in 55 NSCLC tissue samples. N Schematic showing that HHIPL2 regulates Sonic Hedgehog signaling and NSCLC progression by HNF1A. Data in ( A , B , D – H , I – K ) are presented as the mean ± SD. Statistical significance was assessed by a two-sided Student’s t test ( B ), a one-way ANOVA ( A , D – F , H , I , K ), a two-way ANOVA( J ), and a Spearman’s rank correlation coefficient analysis ( M ). * P < 0.05, ** P < 0.01, *** P < 0.001. Experiments ( A , B , D – K ) were repeated at least three times.

Article Snippet: Intriguingly, data from The Human Protein Atlas online database showed that mRNA expression of HHIPL2 was upregulated in the majority of human cancer cell lines compared to non-cancerous cell lines (Fig. ), indicating that higher expression of HHIPL2 is widespread in a pan-cancer manner.

Techniques: Luciferase, Activity Assay, Knockdown, Over Expression, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Migration, Expressing, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: HHIPL2 positively governs Hedgehog signaling to accelerate non-small cell lung cancer progression via enhancing HNRNPC-mediated HNF1A mRNA stabilization

doi: 10.1038/s41419-025-08331-3

Figure Lengend Snippet: A Tandem affinity purification-mass spectrometry detection of HHIPL2-interacting proteins (obtained from FLAG-beads pull down) after A549 cells were transfected with HHIPL2-FLAG for 48 h. B Cell lysates of A549 cells were immunoprecipitated with IgG or HNRNPC antibodies, and immunoblot assays were performed using HHIPL2 and HNRNPC antibodies. C Schematic diagram of HNRNPC and the domain-deleted constructs. RRM, RNA recognition motif; NLS, nuclear localization signal. D Plasmids containing WT, RRM, ΔRRM, and ΔNLS of HNRNPC were co-expressed with HHIPL2-MYC in HEK293T cells. Lysates were immunoprecipitated with FLAG beads. E Best predicted pose of HHIPL2 and HNRNPC, with a prominent ZDOCK score of 1731.296. Active residues forming hydrogen bonds were indicated. F Cellular fractionation analysis of A549 and H157 cells overexpressing HHIPL2-FLAG. Cell fractions were analyzed by Western blot. C cytosol, N nucleus. G Immunofluorescence (IF) staining for endogenous HNRNPC and FLAG in A549 and H157 cells overexpressing FLAG-tagged HHIPL2. Nuclei were stained with DAPI. Scale bar, 10 μm. H Relative RT-qPCR analysis of the mRNA levels of HNF1A in A549 and H157 cells with or without HNRNPC knockdown ( n = 3 in each group). I Knockdown of HNRNPC in A549 and H157 cells. Cell lysates were analyzed by western blotting with antibodies against HNRNPC, HNF1A, and GAPDH. J Binding of HNRNPC protein with HNF1A mRNA was determined by RNA immunoprecipitation (RIP) with FLAG beads in A549 cells and H157 cells overexpressing HNRNPC-FLAG. HNF1A mRNA enrichment was measured by RT-PCR analyses using the relevant primers for HNF1A . K Binding of HNF1A mRNA with HNRNPC protein was determined by RNA pull-down with HNF1A-probe in A549 and H157 cells. HNRNPC protein enrichment was measured by western blotting, and HNF1A mRNA enrichment was measured by RT-PCR analyses using the relevant primers for HNF1A . L , M The half-life of HNF1A mRNA was examined by RT-qPCR in A549 and H157 cells after HNRNPC knockdown and actinomycin D treatment (2 μg/mL) for the indicated times ( n = 3 in each group). N Binding of HNRNPC protein with HNF1A mRNA was determined by RNA immunoprecipitation (RIP) with FLAG beads in A549 and H157 cells overexpressing HNRNPC-FLAG and HHIPL2-GFP. HNF1A mRNA enrichment was measured by RT-PCR analyses using the relevant primers for HNF1A . O Schematic showing that HNRNPC controls HNF1A mRNA stability by HHIPL2-mediated HNRNPC nucleo-cytoplasmic translocation. Data in ( H , L , M ) are presented as the mean ± SD. Statistical significance was assessed by a one-way ANOVA ( H ) and a two-way ANOVA( L , M ). *** P < 0.001. Experiments ( B , D – N ) were repeated at least three times.

Article Snippet: Intriguingly, data from The Human Protein Atlas online database showed that mRNA expression of HHIPL2 was upregulated in the majority of human cancer cell lines compared to non-cancerous cell lines (Fig. ), indicating that higher expression of HHIPL2 is widespread in a pan-cancer manner.

Techniques: Affinity Purification, Mass Spectrometry, Transfection, Immunoprecipitation, Western Blot, Construct, Cell Fractionation, Immunofluorescence, Staining, Quantitative RT-PCR, Knockdown, Binding Assay, RNA Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction, Protein Enrichment, Translocation Assay

Journal: Cell Death & Disease

Article Title: HHIPL2 positively governs Hedgehog signaling to accelerate non-small cell lung cancer progression via enhancing HNRNPC-mediated HNF1A mRNA stabilization

doi: 10.1038/s41419-025-08331-3

Figure Lengend Snippet: Effects of HHIPL2 overexpression with or without HNRNPC knockdown in A549 ( A ) and H157 cells ( B ) using GLI luciferase assays. The GLI luciferase activity is normalized to Renilla ( n = 3 in each group). Relative RT-qPCR analysis of the mRNA levels of the genes related to the Sonic Hedgehog pathway in A549 ( C ) and H157 cells ( D ) with HHIPL2 overexpression and HNRNPC knockdown ( n = 3 in each group). E Overexpression of HHIPL2 in A549 and H157 cells with or without HNRNPC knockdown. Cell lysates were analyzed by western blotting with the indicated antibodies. F Effects of HHIPL2 overexpression with or without HNRNPC knockdown in A549 and H157 cells using Colony formation assays. ImageJ was used to perform quantitative analysis ( n = 3 in each group). Effects of HHIPL2 overexpression with or without HNRNPC knockdown in A549 ( G ) and H157 cells ( H ) using CCK-8 assays ( n = 3 in each group). Effects of HHIPL2 overexpression with or without HNRNPC knockdown on migration and invasion in A549 ( I ) and H157 cells ( J ) using transwell assays and ImageJ was used to perform quantitative analysis ( n = 4 in each group). Scale bar, 100 μm. K Schematic showing that HHIPL2 regulates Sonic Hedgehog signaling and NSCLC progression by HNRNPC. Data in ( A – D , F – J ) are presented as the mean ± SD. Statistical significance was assessed by a one-way ANOVA ( A – D , F , I , J ) and a two-way ANOVA( G , H ). * P < 0.05, ** P < 0.01, *** P < 0.001. Experiments ( A – J ) were repeated at least three times.

Article Snippet: Intriguingly, data from The Human Protein Atlas online database showed that mRNA expression of HHIPL2 was upregulated in the majority of human cancer cell lines compared to non-cancerous cell lines (Fig. ), indicating that higher expression of HHIPL2 is widespread in a pan-cancer manner.

Techniques: Over Expression, Knockdown, Luciferase, Activity Assay, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Migration

Journal: Cell Death & Disease

Article Title: HHIPL2 positively governs Hedgehog signaling to accelerate non-small cell lung cancer progression via enhancing HNRNPC-mediated HNF1A mRNA stabilization

doi: 10.1038/s41419-025-08331-3

Figure Lengend Snippet: A Overexpression of HHIPL2 in A549 and H157 cells in the presence or absence of triptolide (TPL) (50 nM) for 24 h. Cell lysates were analyzed by Western blot. B , C Cell viability was detected by the CCK-8 method, and the dose-response curves of triptolide (TPL) in HHIPL2-overexpression and control cells after 24 h of TPL (0, 2, 4, 8, 16, 32, 64, 128, 256, 512, and 1024 nM) treatment were plotted. The IC 50 value was calculated according to the fitting curves. D Colony formation assays of A549 and H157 cells overexpressing HHIPL2 after treatment with DMSO and triptolide (TPL) (50 nM). ImageJ was used to perform quantitative analysis ( n = 3 in each group). Effects of HHIPL2 overexpression in the presence or absence of triptolide (TPL) (50 nM) on migration and invasion in A549 ( E ) and H157 cells ( F ) using transwell assays, and ImageJ was used to perform quantitative analysis ( n = 4 in each group). Scale bar, 100 μm. BALB/c nude mice were injected subcutaneously with stably transduced A549-Control and A549-HHIPL2 cells. After 27 days, triptolide (TPL) (0.1 mg/kg) was intraperitoneally injected at intervals of 2 days. After 42 days, the mice were then euthanized. The transplanted tumors were removed and photographed ( G ), Tumors were isolated, and the weight ( H ) and volumes ( I ) were measured ( n = 7 per group). Representative images of lung metastasis models in nude mice after tail injection of A549 cells with HHIPL2 overexpression and triptolide (TPL) (0.1 mg/kg) treatment ( J ) and quantification of pulmonary metastatic nodules ( K ) ( n = 5 per group). Representative H&E staining in lung sections ( L ). Data in ( D – F , H , I , K ) are presented as the mean ± SD. Statistical significance was assessed by a one-way ANOVA ( D – F , H , K ) and a two-way ANOVA ( I ). * P < 0.05, ** P < 0.01, *** P < 0.001. Experiments ( A – F ) were repeated at least three times.

Article Snippet: Intriguingly, data from The Human Protein Atlas online database showed that mRNA expression of HHIPL2 was upregulated in the majority of human cancer cell lines compared to non-cancerous cell lines (Fig. ), indicating that higher expression of HHIPL2 is widespread in a pan-cancer manner.

Techniques: Over Expression, Western Blot, CCK-8 Assay, Control, Migration, Injection, Stable Transfection, Isolation, Staining

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: Pan-Cancer Analysis of CD26 expression. A CD26 expression in multiple tissues (HPA database). B Types of cancers with high expression ( p < 0.05) of CD26 (GEPIA2 database). ESCA, Esophageal Carcinoma; KIRP, Kidney Renal Papillary Cell Carcinoma; LAML, Acute Myeloid Leukemia; LIHC, Liver Hepatocellular Carcinoma; LUAD, Lung Adenocarcinoma; PAAD, Pancreatic adenocarcinoma; PRAD, Prostate Adenocarcinoma; STAD, Stomach Adenocarcinoma; THCA, Thyroid Carcinoma; THYM, Thymoma

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Expressing

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: The Association between CD26 Expression and WHO cancer stages. A Pan-cancer. B BRCA. * p < 0.05. CESC, Cervical Squamous Cell Carcinoma and Endocervical Adenocarcinoma; LUAD, Lung Adenocarcinoma; COAD, Colon Adenocarcinoma; COADREAD, Colon adenocarcinoma/Rectum adenocarcinoma Esophageal carcinoma; BRCA, Breast Invasive Carcinoma; ESCA, Esophageal Carcinoma; STES, Stomach and Esophageal carcinoma; KIRP, Kidney Renal Papillary Cell Carcinoma; KIPAN, Pan-kidney cohort (KICH + KIRC + KIRP); STAD, Stomach Adenocarcinoma; UCEC, Uterine Corpus Endometrial Carcinoma; HNSC, Head and Neck Squamous Cell Carcinoma; KIRC, Kidney Renal Clear Cell Carcinoma; LUSC, Lung Squamous Cell Carcinoma; THYM, Thymoma; LIHC, Liver Hepatocellular Carcinoma; THCA, Thyroid Carcinoma; MESO, Mesothelioma; READ, Rectum Adenocarcinoma; PAAD, Pancreatic adenocarcinoma; OV, Ovarian Serous Cystadenocarcinoma; TGCT, Testicular Germ Cell Tumors; SKCM, Skin Cutaneous Melanoma; UVM, Uveal Melanoma; UCS, Uterine Carcinosarcoma; BLCA, Bladder Urothelial Carcinoma; ACC, Adrenocortical Carcinoma; KICH, Kidney Chromophobe; CHOL, Cholangiocarcinoma; DLBC, Lymphoid Neoplasm Diffuse Large B-cell Lymphoma

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Expressing

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: K-M analysis of the association between CD26 expression and OS in different cancers. A Cancers with high CD26 expression for good prognosis. B Cancers with high CD26 expression for poor prognosis. BRCA, Breast Invasive Carcinoma; LAML, Acute Myeloid Leukemia; LUSC, Lung Squamous Cell Carcinoma; PRAD, Prostate Adenocarcinoma; STAD, Stomach Adenocarcinoma; KIRP, Kidney Renal Papillary Cell Carcinoma; LUAD, Lung Adenocarcinoma; SKCM, Skin Cutaneous Melanoma; THCA, Thyroid Carcinoma

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Expressing

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: CD26 expression and the effect of CD26 inhibitor Alogliptin on BRCA cell lines MCF-7 and MCF-10A. A RT-PCR validation of CD26 expression. B - C Western blot analysis to verify CD26 expression in MCF-7 and MCF-10A cells. D - E Western blot analysis to measure functional inhibition of CD26 in MCF-7 and MCF-10A cells. F IC50 assay to analyze the effects of Alogliptin on MCF-7 and MCF-10A cells. G - H Transwell assay to analyze the impact of Alogliptin on MCF-7 cells migration (Scale bar: 100 μm). I ELISA assay to analyze the impact of Alogliptin on MMP9 level of MCF-7 cells. * p < 0.05. IC50: Median Inhibition Concentration, ELISA: Enzyme-Linked Immunosorbent Assay; MMP9: Matrix metallopeptidase 9

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Western Blot, Functional Assay, Inhibition, Transwell Assay, Migration, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the top 100 genes co-expressed with CD26 and 10 genes with the proteins bound to CD26. A GO analysis. B KEGG analysis

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques:

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: Correlation analysis between CD26 expression and immune cell infiltration levels in pan-cancer. THYM, Thymoma; SKCM, Skin Cutaneous Melanoma; KIRC, Kidney Renal Clear Cell Carcinoma; LGG, Brain Lower Grade Glioma; PCPG: Pheochromocytoma and Paraganglioma; THCA, Thyroid Carcinoma; BRCA, Breast Invasive Carcinoma; LUSC, Lung Squamous Cell Carcinoma; LIHC, Liver Hepatocellular Carcinoma; KICH, Kidney Chromophobe; LUAD, Lung Adenocarcinoma; TGCT, Testicular Germ Cell Tumors; UCS, Uterine Carcinosarcoma; GBMLGG, Glioma; KIPAN, Pan-kidney cohort (KICH + KIRC + KIRP); READ, Rectum Adenocarcinoma; COADREAD, Colon adenocarcinoma/Rectum adenocarcinoma Esophageal carcinoma; STAD, Stomach Adenocarcinoma; PRAD: Prostate Adenocarcinoma; CESC, Cervical Squamous Cell Carcinoma and Endocervical Adenocarcinoma; PAAD, Pancreatic adenocarcinoma; KIRP, Kidney Renal Papillary Cell Carcinoma; STES, Stomach and Esophageal carcinoma; COAD, Colon Adenocarcinoma; ACC, Adrenocortical Carcinoma; SARC, Sarcoma; GBM, Glioblastoma Multiforme; HNCA, Head and Neck Squamous Cell Carcinoma; BLCA, Bladder Urothelial Carcinoma; UVM, Uveal Melanoma; CHOL, Cholangiocarcinoma; MESO, Mesothelioma; OV, Ovarian Serous Cystadenocarcinoma; ESCA, Esophageal Carcinoma; UCEC, Uterine Corpus Endometrial Carcinoma; DLBC, Lymphoid Neoplasm Diffuse Large B-cell Lymphoma

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Expressing

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: Correlation analysis between CD26 expression and immune checkpoints in pan-cancer. LUAD, Lung Adenocarcinoma; WT, High-Risk Wilms Tumor; PAAD, Pancreatic adenocarcinoma; READ, Rectum Adenocarcinoma; HNSC, Head and Neck Squamous Cell Carcinoma; OV, Ovarian Serous Cystadenocarcinoma; KICH, Kidney Chromophobe; KIPAN, Pan-kidney cohort (KICH + KIRC + KIRP); KIRC, Kidney Renal Clear Cell Carcinoma; CHOL, Cholangiocarcinoma; UCS, Uterine Carcinosarcoma; COAD, Colon Adenocarcinoma; COADREAD, Colon adenocarcinoma/Rectum adenocarcinoma Esophageal carcinoma; SKCM, Skin Cutaneous Melanoma; UCEC, Uterine Corpus Endometrial Carcinoma; THCA, Thyroid Carcinoma; GBM, Glioblastoma Multiforme; LGG, Brain Lower Grade Glioma; SARC, Sarcoma; PCPG: Pheochromocytoma and Paraganglioma; BRCA, Breast Invasive Carcinoma; LUSC, Lung Squamous Cell Carcinoma; ALL, Acute Lymphoblastic Leukemia; LAML: Acute Myeloid Leukemia; DLBC, Lymphoid Neoplasm Diffuse Large B-cell Lymphoma; NB, Neuroblastoma; BLCA, Bladder Urothelial Carcinoma; UVM, Uveal Melanoma; THYM, Thymoma; TGCT, Testicular Germ Cell Tumors; KIRP, Kidney Renal Papillary Cell Carcinoma; ACC, Adrenocortical Carcinoma; CESC, Cervical Squamous Cell Carcinoma and Endocervical Adenocarcinoma; STAD, Stomach Adenocarcinoma; STES, Stomach and Esophageal carcinoma; ESCA, Esophageal Carcinoma; MESO, Mesothelioma; LIHC, Liver Hepatocellular Carcinoma; PRAD: Prostate Adenocarcinoma

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Expressing, Wilms Tumor Assay

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: CD26 Gene mutation information in pan-cancer. TMB: Tumor Mutational Burden, MSI: Microsatellite Instability; NEO: Neoantigen. GBMLGG, Glioma; LGG, Brain Lower Grade Glioma; CESC, Cervical Squamous Cell Carcinoma and Endocervical Adenocarcinoma; LUAD, Lung Adenocarcinoma; COAD, Colon Adenocarcinoma; COADREAD, Colon adenocarcinoma/Rectum adenocarcinoma Esophageal carcinoma; LAML: Acute Myeloid Leukemia; BRCA, Breast Invasive Carcinoma; ESCA, Esophageal Carcinoma; STES, Stomach and Esophageal carcinoma; SARC, Sarcoma; KIRP, Kidney Renal Papillary Cell Carcinoma; KIPAN, Pan-kidney cohort (KICH + KIRC + KIRP); STAD, Stomach Adenocarcinoma; UCEC, Uterine Corpus Endometrial Carcinoma; HNSC, Head and Neck Squamous Cell Carcinoma; KIRC, Kidney Renal Clear Cell Carcinoma; LUSC, Lung Squamous Cell Carcinoma; THYM, Thymoma; LIHC, Liver Hepatocellular Carcinoma; PAAD, Pancreatic adenocarcinoma; OV, Ovarian Serous Cystadenocarcinoma; TGCT, Testicular Germ Cell Tumors; SKCM, Skin Cutaneous Melanoma; UCS, Uterine Carcinosarcoma; BLCA, Bladder Urothelial Carcinoma

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Mutagenesis

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: Spearman correlation between CD26 expression and ( A )Tumor Mutational Burden (TMB), ( B ) microsatellite instability (MSI) and ( C ) Neoantigen (NEO) in pan-cancer. THYM, Thymoma; MESO, Mesothelioma; PRAD: Prostate Adenocarcinoma; UVM, Uveal Melanoma; KICH, Kidney Chromophobe; STAD, Stomach Adenocarcinoma; DLBC, Lymphoid Neoplasm Diffuse Large B-cell Lymphoma; LUAD, Lung Adenocarcinoma; CESC, Cervical Squamous Cell Carcinoma and Endocervical Adenocarcinoma; KIRC, Kidney Renal Clear Cell Carcinoma; STES, Stomach and Esophageal carcinoma; LAML: Acute Myeloid Leukemia; PAAD, Pancreatic adenocarcinoma; ACC, Adrenocortical Carcinoma; LGG, Brain Lower Grade Glioma; BRCA, Breast Invasive Carcinoma; BLCA, Bladder Urothelial Carcinoma; GBM, Glioblastoma Multiforme; THCA, Thyroid Carcinoma; HNSC, Head and Neck Squamous Cell Carcinoma; GBMLGG, Glioma; LUSC, Lung Squamous Cell Carcinoma; TGCT, Testicular Germ Cell Tumors; KIPAN, Pan-kidney cohort (KICH + KIRC + KIRP); READ, Rectum Adenocarcinoma; LIHC, Liver Hepatocellular Carcinoma; PCPG: Pheochromocytoma and Paraganglioma; UCS, Uterine Carcinosarcoma; SARC, Sarcoma; SKCM, Skin Cutaneous Melanoma; COADREAD, Colon adenocarcinoma/Rectum adenocarcinoma Esophageal carcinoma; COAD, Colon Adenocarcinoma; CHOL, Cholangiocarcinoma; UCEC, Uterine Corpus Endometrial Carcinoma; OV, Ovarian Serous Cystadenocarcinoma; KIRP, Kidney Renal Papillary Cell Carcinoma; ESCA, Esophageal Carcinoma

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Expressing

Journal: BMC Cancer

Article Title: The role of CD26 in breast cancer and its pan-cancer analysis

doi: 10.1186/s12885-026-15656-5

Figure Lengend Snippet: The potential mechanisms by which CD26 may be involved in tumors. M1, M1 macrophage; M2, M2 macrophage; ADA, Adenosine deaminase; FAPα, Fibroblast activation protein α; MMPs, Matrix metalloproteinases; EGF, Epidermal growth factor; ERK, Extracellular-signal regulated kinase; bFGF, Basic fibroblast growth factor; MEK, Mitogen-activated protein kinase

Article Snippet: The CD26 (ENSG00000197635) mRNA expression map in tissues was constructed by the Human Protein Atlas (HPA) ( https://www.proteinatlas.org/ ) database (version: 22.0, Release date: 2022.12.07).

Techniques: Activation Assay

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: CLDN3 expression in normal and cancer tissues. (A) CLDN3 expression in human tissues based on HPA RNA‐seq data. (B) Protein expression levels of CLDN3 in human tissues, data is from the HPA database. (C) CLDN3 expression in different cancers from TIMER2. (D) Expression of CLDN3 across cancers form CPTAC samples. (E) CLDN3 expression in COAD, THYM, UCS, OV, READ, SCKM and SARC (data from GEPIA2). (F) The IHC images of CLDN3 in normal and cancer tissues of LIHC, COAD and READ. (G) Expression levels of CLDN3 total protein in lung adenocarcinoma, UCEC, ovarian cancer, colon cancer and breast cancer. (H) The subcellular location of CLDN3 by indirect immunofluorescence microscopy.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Expressing, RNA Sequencing, Immunofluorescence, Microscopy

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Diagnosis and survival analysis of CLDN3 in cancers. (A) Associations between CLDN3 expression and overall survival across human cancers from TISIDB. (B) Associations between CLDN3 expression and overall survival across LUSC and MESO from GEPIA2. (C) Effects of CLDN3 expression on overall survival in multiple cancer types (data from KM‐Plotter). (D) ROC analysis of CLDN3 in different cancers.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Biomarker Discovery, Expressing

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Epigenetic features of CLDN3 in cancers. (A) Alteration frequencies of CLDN3 across different tumours from cBioPortal. (B) General mutation counts of CLDN3 in various TCGA cancer types from cBioPortal. (C) Mutation types and sites of CLDN3 from cBioPortal. (D) CNV (copy number variation) percentage of CLDN3 in each cancer type. (E) Correlations of CNV with mRNA expression of CLDN3. (F) Correlations of CLDN3 mRNA expression with MSI and TMB in various cancers.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Mutagenesis, Expressing

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Methylation analysis of CLDN3 across various cancer types. (A) Methylation differences between tumour and normal samples of CLDN3 (data from GSCA). (B) Correlations between methylation and mRNA expression of CLDN3 in the specific cancers (data from GSCA). (C) Survival differences between high and low methylation of CLDN3 in specific cancers (data from GSCA).

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Methylation, Expressing

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Interactions between tumour‐immune system and CLDN3. (A) Correlations of CLDN3 mRNA expression with TILs, immunoinhibitors, immunostimulators, MHC, chemokines and receptors across human cancers (data from TISIDB). (B) Expression and mutation differences for CLDN3 between responders and non‐responders (data from TISIDB).

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Expressing, Mutagenesis

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Interactions between CLDN3 and immune infiltration. Positive correlation between CLDN3 expression and immune infiltration of T cell regulatory (Tregs) in all (A) and specific cancers (B). Negative correlation between CLDN3 expression and immune infiltration of MAST cells in all (C) and specific cancers (D).

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Expressing

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Identification of CLDN3‐related genes and their associated biological functions. (A) protein–protein interaction (PPI) analysis for CLDN3‐interacting proteins, data from STRING. Pathway and process enrichment analysis of genes coding CLDN3‐interacting proteins from KEGG (B) and Metascape (C). Corresponding heat maps of 10 CLDN3‐related genes in specific cancer types, (D) 10 genes from GEPIA2; (E) 10 genes from PPI. (F) Wayne diagrams of intersection analyses of CLDN3‐correlated and inter‐acting genes (11 counts). (G) Cancer related pathway analysis of the 11 genes. (H) Pathway analysis of the 11 genes.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques:

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Validation of CLDN3 expression and function in CRC cells. (A) Correlations of CLDN3 mRNA expression with cells function in cancers. (B) Single‐cell analysis of CLDN3 in CRC_GSE146771‐Smartseq2 dataset. (C) Analysis of CLDN3 expression in different CRC dataset. (D) Analysis of CLDN3 expression in different cells by using TISCH2 database data from CRC_GSE146771‐Smartseq2. (E) Promoter methylation analysis of CLDN3 in individual cancer stages and tumour grades. Relative (F) protein and (G) mRNA expression of CLDN3 in stably CLDN3‐knockdown and ‐overexpression CRC cell lines. Cell proliferation assays for CLDN3‐knockdown and ‐overexpression CRC cells using the (H) CCK8 assay, (I) EdU staining assay and (J) plate colony formation assay. (K) Wound healing scratch assays using CLDN3‐knockdown and ‐overexpression CRC cells. (L) Cell cycle analysis for the CRC cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Biomarker Discovery, Expressing, Single-cell Analysis, Methylation, Stable Transfection, Knockdown, Over Expression, CCK-8 Assay, Staining, Colony Assay, Cell Cycle Assay

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: CLDN3 enhances the chemoresistance to 5‐FU in CRC cells. (A) Correlation between drug sensitivity and CLDN3 expression, data from CellMiner. Correlations between CLDN3 expression with the sensitivity of CTRP drugs (B) and GDSC drugs (C) in pan‐cancer (data from GSCA). (D) IC50 of 5‐fluorouracil (5‐FU) for HCT116 and SW480 cells. (E) Apoptosis of 5‐FU‐treated CRC cells with CLDN3 stable overexpression or CLDN3 knockdown as indicated. (F) Identifying the CLDN3‐regulated signalling pathways via human phospho‐kinase array. (G) Under conditions of P38 pathway inhibition, apoptosis of 5‐FU‐treated CLDN3 stable overexpression CRC cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Expressing, Over Expression, Knockdown, Inhibition

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Trim28 is an interacting protein of CLDN3. (A) Silver staining assays and mass spectrometry to investigate CLDN3‐associated proteins in colorectal cancer cells. (B) KEGG pathways enrichment analysis of CLDN3‐associated proteins. Correlation between CLDN3 and TRIM28 expression across multiple malignancies, data from GEPIA2 (C) and data from TIMER2.0 (D).

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Silver Staining, Mass Spectrometry, Expressing

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: Trim28 catalysed the SUMOylation of CLDN3 to regulate its stability. (A) Protein‐protein interaction between endogenous CLDN3 and TRIM28. (B) Protein‐protein interaction between exogenous HA‐CLDN3 and GFP‐TRIM28. (C) Intracellular localisation of CLDN3 and TRIM28 in CRC cells. (D) Co‐IPs as indicated demonstrating that TRIM28 expression promotes the SUMOylation of endogenous CLDN3. (E) TRIM28 expression promotes the SUMOylation of exogenous CLDN3. (F) Downregulated TRIM28 expression in HCT116 cells using RNA interference. (G) Degradation of CLDN3 in the CRC cells with TRIM28 knockdown.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Expressing, Knockdown

Journal: IET Systems Biology

Article Title: Pan‐Cancer Analysis of CLDN3 and Its Contribution to 5‐FU Resistance in Colorectal Cancer

doi: 10.1049/syb2.70059

Figure Lengend Snippet: SUMOylation of CLDN3 at Lys156 regulates its protein stability and promotes CRC progression. (A) Prediction of potential SUMOylation sites in the CLDN3 protein. (B) Analysis of SUMOylation in HEK293T cells with wild‐type CLDN3, CLDN3 K156A and CLDN3 K190A . (C) Detection of protein stability of CLDN3 WT or CLDN3 K156A using immunoblotting assays. (D) Colony‐forming capacity of CRC cells with CLDN3 WT or CLDN3 K156A overexpression as indicated. (E) Migration and invasion of CRC cells with CLDN3 WT or CLDN3 K156A overexpression.

Article Snippet: CLDN3 mRNA expression in normal tissues was obtained from the human protein atlas (HPA) ( http://www.proteinatlas.org ) [ ], based on RNA‐seq data.

Techniques: Western Blot, Over Expression, Migration

Journal: bioRxiv

Article Title: High prevalence of CNS-directed autoantibodies in patients with schizophrenia

doi: 10.64898/2026.05.04.722731

Figure Lengend Snippet: A. Receiver Operator Classification (ROC) curve of the REAP-Lasso model for patient vs. control discrimination (AUC = 0.884). B. The number of common autoantibody (AAb) reactivities per individual by group (control, n=971; schizophrenia, n = 352). Common reactivities defined as present in >1% of the control cohort. Significance was assessed using unpaired two-sided Wilcoxon test. For the box plots, the central lines indicate the group median values, the top and bottom lines indicate the 75th and 25th percentiles, respectively, the whiskers represent 1.5× the interquartile range. C. The number of low-frequency autoantibody (AAb) reactivities per individual by group (control, n=971; schizophrenia, n = 352). Low-frequency reactivities defined as present in <= 1% of the control cohort. Significance was assessed using unpaired two-sided Wilcoxon test. For the box plots, the central lines indicate the group median values, the top and bottom lines indicate the 75th and 25th percentiles, respectively, the whiskers represent 1.5× the interquartile range. Low-freq = Low-frequency D. The number of autoantibody (AAb) reactivities against each tissue category per individual by group (control, n = 971; schizophrenia = 352). Tissue categories are composed of REAP reactivities bucketed by human protein atlas mRNA expression data. Significance was assessed by unpaired two-sided Wilcoxon with correction for multiple hypotheses by Benjamini-Hochberg. *** ∼ <0.001, ** ∼ <0.01, * ∼ <0.05. E, F. Forest plots depicting the top tissues (E) or single cell types (F) (y-axis) ranked by their estimated contribution to the overall schizophrenia vs. control increase in total autoantibodies (AAbs), with the x-axis reporting percentage of overall schizophrenia increase explained. Tissue or single cell categories are composed of REAP reactivities bucketed by human protein atlas mRNA expression data. Because a given protein may belong to more than one tissue class, percentages need not sum to 100. Closed circles (●) represent the observed percent contribution. Open circles (○) indicate the expected contribution under a REAP-library baseline. Horizontal whiskers denote 95% confidence intervals from a cohort-/group-stratified bootstrap that resamples subjects within schizophrenia and control groups (R = 4000 replicates).

Article Snippet: To assess whether specific tissues or cell types are preferentially targeted in schizophrenia, we annotated REAP antigens by tissue and cell-type expression using Human Protein Atlas (HPA) mRNA data.

Techniques: Control, Expressing, Single Cell

Journal: bioRxiv

Article Title: High prevalence of CNS-directed autoantibodies in patients with schizophrenia

doi: 10.64898/2026.05.04.722731

Figure Lengend Snippet: A. The number of autoantibody (AAb) reactivities against each tissue category per individual by group (control, n = 971; schizophrenia = 352). Single cell categories are composed of REAP reactivities bucketed by human protein atlas mRNA expression data. Significance was assessed by unpaired two-sided Wilcoxon with correction for multiple hypotheses by Benjamini-Hochberg. *** ∼ <0.001, ** ∼ <0.01, * ∼ <0.05.

Article Snippet: To assess whether specific tissues or cell types are preferentially targeted in schizophrenia, we annotated REAP antigens by tissue and cell-type expression using Human Protein Atlas (HPA) mRNA data.

Techniques: Control, Single Cell, Expressing

Journal: bioRxiv

Article Title: High prevalence of CNS-directed autoantibodies in patients with schizophrenia

doi: 10.64898/2026.05.04.722731

Figure Lengend Snippet: A. Conceptual figure for depiction of autoantibody odds ratio for schizophrenia status. B. Volcano plot depicting all autoantibody reactivities detected. Reactivities that are significantly enriched in schizophrenia (q-value <0.05, odds ratio (OR) >2.72) and target proteins with elevated expression in the brain and inhibitory and excitatory neurons by human protein atlas mRNA expression are bolded and colored. Color indicates functional annotation. Each dot represents one autoantibody reactivity. Vertical dashed lines indicate OR threshold +/-2.72. Horizontal dashed line indicates q-value threshold of 0.05. C. Conceptual figure for depiction of autoantibody odds ratio for schizophrenia status and AAb frequency in schizophrenia. D,E,F. Frequency plot depicting all autoantibody reactivities detected, with proteins belonging to the ion channel and Ca+2 excitability group (D) , synaptic function and synaptic plasticity groups (E) , and neuromodulatory GPCR (F) from figure B bolded and colored. Horizontal dashed lines indicate OR threshold of +/-2.72. Each dot represents one reactivity. G. Volcano plot depicting all anti-pathogen reactivities detected. Reactivities that are significantly enriched in schizophrenia (q-value <0.05, odds ratio (OR) >2.72) are bolded and colored. Color indicates pathogen class annotation. Each dot represents one reactivity. Vertical dashed lines indicate OR threshold +/-2.72. Horizontal dashed line indicates q-value threshold of 0.05

Article Snippet: To assess whether specific tissues or cell types are preferentially targeted in schizophrenia, we annotated REAP antigens by tissue and cell-type expression using Human Protein Atlas (HPA) mRNA data.

Techniques: Expressing, Functional Assay

Journal: bioRxiv

Article Title: High prevalence of CNS-directed autoantibodies in patients with schizophrenia

doi: 10.64898/2026.05.04.722731

Figure Lengend Snippet: A. Volcano plot depicting all autoantibody reactivities detected, with reactivities that are significantly enriched in schizophrenia (q-value <0.05, odds ratio (OR) >2.71) and target proteins elevated in brain pericytes, endothelial cells, astrocytes, vascular associated smooth muscle, or choroid plexus epithelial cells (by HPA mRNA expression data) bolded and colored. Each dot represents one autoantibody reactivity. Vertical dashed lines indicate OR threshold +/-2.71. Horizontal dashed line indicates q-value threshold of 0.05. B. Graphic representation of the MIMETAS chip used for the 3D human BBB model with flow. C. Diagram illustrating the creation of the 3D human BBB model and experimental scheme for the tracer permeability assay. D. Graphic representation of the experimental scheme for the TEER measurement with the OrganoTEER instrument. E. Graph of the TEER measurements over time in the 3D human BBB model under three conditions. Each dot represents one measurement. ** p<0.01, **** p<0.001. F. Representative images of tracer leakage from the blood (left) into the brain (right) compartment of the 3D BBB model. Biocytin is in green and Dextran is in red. There is increased tracer in the brain compartment with the BBB positive sera from Schizophrenia patients. G,H. Plots of the normalized signal of biocytin (G) and 70 KDa Dextran (H) permeability measurements at 30 minutes after tracer application. Significance was assessed using one way ANOVA (p=3.2E-9, 2.6E-12) followed by Tukey’s post hoc test. For the box plots, the central lines indicate the group median values, the top and bottom lines indicate the 75th and 25th percentiles, respectively, the whiskers represent 1.5× the interquartile range. BBB = Blood brain barrier. I. Number of autoantibodies (AAb) per individual (ID) targeting proteins with elevated expression in the brain by human protein atlas mRNA expression data. N=328 for BBB-, 24 for BBB+. Significance was assessed using unpaired Wilcoxon. For the box plots, the central lines indicate the group median values, the top and bottom lines indicate the 75th and 25th percentiles, respectively, the whiskers represent 1.5× the interquartile range. BBB = Blood-brain barrier.

Article Snippet: To assess whether specific tissues or cell types are preferentially targeted in schizophrenia, we annotated REAP antigens by tissue and cell-type expression using Human Protein Atlas (HPA) mRNA data.

Techniques: Expressing, Permeability

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: Conceptual model of ENOX2-SIRT1-mediated regulation of cancer cell functions. ENOX2/tNOX catalyzes the oxidation of NADH to NAD + , leading to activation of the NAD + -dependent deacetylase SIRT1. Activated SIRT1 deacetylates downstream proteins and transcription factors, influencing cancer cell proliferation, migration, invasion, and cell death.

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Activation Assay, Histone Deacetylase Assay, Migration

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: Expression of ENOX2 in head and neck cancer. ( A , B ). Differential expression of ENOX2 in HNSCC and adjacent normal tissues, analyzed from the TCGA-HNSC (DOI: 10.1038/nature14129) ( A ) and CPTAC ( B ) datasets using UALCAN ( https://ualcan.path.uab.edu ). ENOX2 mRNA levels were significantly higher in TCGA-HNSC tumor tissues than in normal tissues (*** p < 0.001), whereas CPTAC proteomic data showed a non-significant trend toward higher ENOX2 protein expression in tumors. ( C ). Analysis of transcriptomic data from the Human Protein Atlas (HPA) based on the CCLE dataset, showing that ENOX2 mRNA expression varies among head and neck cancer cell lines, with relatively higher expression observed in the HSC-3 cell line. Statistical testing ( p -values) was not available from the dataset. ( D ) Heatmap visualization of ENOX2 mRNA expression across multiple cancer types in the HPA dataset, showing that ENOX2 overexpression is not limited to HNSCC. Values are normalized within the dataset and are not directly compared to the housekeeping gene.

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Expressing, Quantitative Proteomics, Over Expression

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: Expression and clinical relevance of ENOX2, SIRT1 , and SOX2 in head and neck cancer datasets. ( A ) TCGA analysis revealed significantly elevated SIRT1 and SOX2 transcript levels in HNSCC tumor tissues compared to normal tissues across all histological grades (* p < 0.05, *** p < 0.001). ( B ) Pearson correlation analysis demonstrated positive correlations between ENOX2 and SIRT1 , as well as between ENOX2 and SOX2 . ( C ) Kaplan–Meier plotter analysis indicated that high mRNA expression levels of ENOX2 and SOX2 were significantly associated with poor overall survival in both female and male patients with head and neck cancer, whereas SIRT1 expression showed a significant association only in the female group (stage III + IV).

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Expressing

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: ENOX2 prompts stem-like properties and proliferation in oral cancer cells. ( A ) Interaction network of ENOX2 and associated proteins generated using the Integrated Interaction Database (IID). ( B ) Co-immunoprecipitation of GST-ENOX2-overexpressing cell lysates with non-immune IgG or antibodies against GST and SOX2, followed by Western blotting with anti-GST or anti-SOX2 antibodies. Whole-cell lysates were also immunoblotted with anti-ENOX2 and anti-SOX2 antibodies; β-actin served as a loading control. Representative images are shown. ( C , D ) Western blot analysis of stemness markers in cells transfected with GST or GST-ENOX2 for 72 h ( C ), or with si-control (scramble RNAi) or si-ENOX2 for 48 h ( D ). β-actin was used as the loading control. ( E ) Real-time monitoring of proliferation in SAS and HSC-3 cells transfected with GST, GST-ENOX2, si-control, or si-ENOX2. After overnight attachment, cells were seeded onto E-plates, and proliferation was continuously measured using the xCELLigence system. Cell index values are presented. ( F ) Pearson correlation analysis of ENOX2 with XIAP and MRPL41 in TCGA tumor tissues using UALCAN ( https://ualcan.path.uab.edu ).

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Generated, Immunoprecipitation, Western Blot, Control, Transfection

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: Sphere formation and stem-like properties of SAS and HSC-3 oral cancer cells. ( A ) Sphere-forming abilities of SAS and HSC-3 cells. Various cell densities of SAS cells (0.5 × 10 3 , 1 × 10 3 , 2 × 10 3 , 3 × 10 3 , 4 × 10 3 , 5 × 10 3 , and 1 × 10 4 ) and HSC-3 cells (1 × 10 4 ) were seeded into 96-well ultra-low attachment plates and cultured in DMEM supplemented with 10% FBS at 37 °C. Representative images (magnification 10×) obtained at different time points are shown. ( B ) Western blot analysis of spheroid cells harvested at day 7, showing expression of ENOX2, SIRT1, and stemness-associated markers. β-Actin served as the loading control. ( C ) Comparison of the intracellular NAD + /NADH ratios between adherent and spheroid SAS cells, as measured using an NADH/NAD quantification kit. Data are presented as mean ± SD from three independent experiments (** p < 0.01).

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Cell Culture, Western Blot, Expressing, Control, Comparison

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: Role of ENOX2 in regulating the stemness of oral cancer cells. ( A ) SAS and HSC-3 cells were transfected with GST or GST-ENOX2 for 72 h, seeded onto poly-HEMA-coated 6-well plates, and cultured under non-adherent conditions. After 7 days, spheroids were harvested, and protein levels of stemness-associated markers were analyzed by Western blotting. ( B ) SAS and HSC-3 cells were transfected with si-control or si-ENOX2 for 48 h, seeded onto poly-HEMA-coated 6-well plates, and cultured under non-adherent conditions. After 7 days, spheroids were harvested and analyzed by Western blotting. β-Actin was used as the loading control.

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Transfection, Cell Culture, Western Blot, Control

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: ENOX2 promotes the growth of oral cancer xenografts in ASID mice. ( A ) SAS cells were transiently transfected with GST or GST-ENOX2. After 72 h, cells were cultured in poly-HEMA–coated plates to form spheres. Following 7 days of sphere formation, spheroids were dissociated into single cells and subcutaneously inoculated into ASID mice at doses of 1 × 10 6 , 2 × 10 4 , or 1 × 10 4 . Tumor growth was assessed by measuring tumor volume. Representative tumor morphology and quantitative analyses are shown. Statistically significant differences between the GST and GST-ENOX2 groups were determined by one-way ANOVA with LSD post hoc test (* p < 0.05, ** p < 0.01, *** p < 0.001). ( B ) Tumor tissues from both groups were homogenized and analyzed by Western blotting for stemness-associated markers. ( C ) Sphere-forming cells transfected with si-control or si-ENOX2 (1 × 10 6 ) were subcutaneously inoculated into ASID mice (n = 3 per group). Tumor growth, measured by assessment of tumor volume, was significantly reduced in the si-ENOX2 group compared with controls (* p < 0.05, *** p < 0.001).

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Transfection, Cell Culture, Western Blot, Control

Journal: Antioxidants

Article Title: ENOX2 (tNOX)–Associated Stemness in Oral Cancer Cells and Its Clinical Correlation in Head and Neck Tumors

doi: 10.3390/antiox15010098

Figure Lengend Snippet: Capsaicin suppresses sphere formation and stemness marker expression in SAS cells. ( A ) SAS cells were cultured in ultra-low attachment plates for 48 h, and then treated with or without capsaicin for 5 days. Representative images of spheres at day 7 are shown. ( B ) Western blot analysis of SAS spheroids treated with or without capsaicin. Protein levels of ENOX2, SIRT1, and stemness-associated markers (SOX2, Nanog, Oct4) were assessed. β-Actin served as a loading control.

Article Snippet: Consistent with these findings, data from the Human Protein Atlas ( https://www.proteinatlas.org ) showed that ENOX2 mRNA expression was elevated in head and neck cancer cell lines, with relatively higher levels observed in HSC-3 cells ( C).

Techniques: Marker, Expressing, Cell Culture, Western Blot, Control

Journal: Scientific Reports

Article Title: Characterization of growth differentiation factor 15 (GDF15) as a neurotropic adipokine permeable to the brain

doi: 10.1038/s41598-025-14846-8

Figure Lengend Snippet: Expression of GDF15 mRNA and secretion of GDF15 protein in 3T3-L1 adipocytes. Panel A Upregulation of GDF15 mRNA expression by qPCR during the differentiation of 3T3-L1 pre-adipocytes (day 0) into mature adipocytes (day 8). The Kruskal-Wallis test was performed. Panel B Secretion of GDF15 protein into supernatants by ELISA is upregulated by insulin under normo- and hyperglycemic conditions. The Kruskal-Wallis test was performed.

Article Snippet: As provided by the Human Protein Atlas ( https://www.proteinatlas.org/ENSG00000130513-GDF15/tissue ), GDF15 mRNA expression is detectable in multiple organs.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Characterization of growth differentiation factor 15 (GDF15) as a neurotropic adipokine permeable to the brain

doi: 10.1038/s41598-025-14846-8

Figure Lengend Snippet: Correlation analysis by scatter plot diagrams. Panel A Positive correlation of serum and CSF GDF15 concentrations. Panel B Correlation of GDF15 concentrations in CSF with age. Panel C Correlation of GDF15 concentrations in serum with age. The Spearman-rho rank correlation test was applied.

Article Snippet: As provided by the Human Protein Atlas ( https://www.proteinatlas.org/ENSG00000130513-GDF15/tissue ), GDF15 mRNA expression is detectable in multiple organs.

Techniques:

Journal: Scientific Reports

Article Title: Characterization of growth differentiation factor 15 (GDF15) as a neurotropic adipokine permeable to the brain

doi: 10.1038/s41598-025-14846-8

Figure Lengend Snippet: Dependence of GDF15 concentrations on gender, obesity and type 2 diabetes. Panel A GDF15 concentrations in CSF are higher in males than in females. Panel B GDF15 concentrations in serum show no gender-specific effect. Panel C GDF15 concentrations in serum are higher in overweight/obese patients. Panel D GDF15 concentrations in CSF are higher in overweight/obese patients. Panel E GDF15 concentrations in serum are higher in type 2 diabetic patients. Panel F GDF15 concentrations in CSF are higher in type 2 diabetic patients. The Mann-Whitney U test was applied.

Article Snippet: As provided by the Human Protein Atlas ( https://www.proteinatlas.org/ENSG00000130513-GDF15/tissue ), GDF15 mRNA expression is detectable in multiple organs.

Techniques: MANN-WHITNEY

Journal: Scientific Reports

Article Title: Characterization of growth differentiation factor 15 (GDF15) as a neurotropic adipokine permeable to the brain

doi: 10.1038/s41598-025-14846-8

Figure Lengend Snippet: Correlation analysis (scatter plot diagrams) of CSF and serum concentrations of GDF15 with serum glucose and HbA 1c levels. Panel A Positive correlation of GDF15 concentrations in CSF with serum glucose. Panel B Positive correlation of GDF15 concentrations in serum with serum glucose. Panel C Positive correlation of GDF15 concentrations in CSF with HbA 1c levels. Panel D Positive correlation of GDF15 concentrations in serum with HbA 1c levels. The Spearman-rho rank correlation test was applied.

Article Snippet: As provided by the Human Protein Atlas ( https://www.proteinatlas.org/ENSG00000130513-GDF15/tissue ), GDF15 mRNA expression is detectable in multiple organs.

Techniques:

Journal: Scientific Reports

Article Title: Characterization of growth differentiation factor 15 (GDF15) as a neurotropic adipokine permeable to the brain

doi: 10.1038/s41598-025-14846-8

Figure Lengend Snippet: GDF15 in neurological disease groups. Panel A GDF15 concentrations in CSF are higher in patients with infectious diseases (and cardiovascular diseases) in the entire study cohort. The Kruskal-Wallis test was performed. Panel B GDF15 concentrations in CSF are higher in patients with infectious diseases when compared to matched controls. The Mann-Whitney U test was applied. Panel C GDF 15 concentrations in serum are not correlated to infectious diseases but only to cardiovascular diseases. Ctrl, control group; CD, vascular diseases; ID, infectious diseases; MS, multiple sclerosis; PC, pseudotumor cerebri. The Kruskal-Wallis test was performed.

Article Snippet: As provided by the Human Protein Atlas ( https://www.proteinatlas.org/ENSG00000130513-GDF15/tissue ), GDF15 mRNA expression is detectable in multiple organs.

Techniques: MANN-WHITNEY, Control

Journal: Scientific Reports

Article Title: Characterization of growth differentiation factor 15 (GDF15) as a neurotropic adipokine permeable to the brain

doi: 10.1038/s41598-025-14846-8

Figure Lengend Snippet: GDF15 concentrations in CSF and their relation to CSF cell count and dysfunction of the blood–brain-barrier (BBB). Panel A GDF15 concentrations in CSF are higher in patients with elevated CSF cell count. The Mann-Whitney U test was applied. Panel B GDF15 concentrations in CSF are higher in patients with dysfunction of BBB. The Kruskal-Wallis test was performed.

Article Snippet: As provided by the Human Protein Atlas ( https://www.proteinatlas.org/ENSG00000130513-GDF15/tissue ), GDF15 mRNA expression is detectable in multiple organs.

Techniques: Cell Counting, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Characterization of growth differentiation factor 15 (GDF15) as a neurotropic adipokine permeable to the brain

doi: 10.1038/s41598-025-14846-8

Figure Lengend Snippet: Clinical procedure and calculation of specific CSF/serum values for GDF15. BBB permeability of GDF15 and its relation to other classical adipokines is given at a glance. The higher the CSF/serum ratio, the higher the grade of permeability to the brain. Due to the percentage of the CSF protein quantity in relation to the quantity in peripheral blood serum, three grades of permeability (1/1000; 1/100; 1/10) can be defined for a better clinical undertanding. The famous, adipocyte-specific, satiety hormone leptin signals to the brain and has a quantitiy in CSF of 1/1000 in relation to peripheral blood serum. BBB, blood–brain-barrier; CSF, cerebrospinal fluid. Created with a BioRender personal account.

Article Snippet: As provided by the Human Protein Atlas ( https://www.proteinatlas.org/ENSG00000130513-GDF15/tissue ), GDF15 mRNA expression is detectable in multiple organs.

Techniques: Permeability

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: A Par6 mRNA levels in various cancers from The Cancer Genome Atlas (TCGA) database ( n = 7932). B , C Determination of Par6 expression in HEB, U87MG, T98G, and GBM1 cell lines at the transcriptional and translational levels ( n = 3 for each group). D Determination of the overexpression and knockdown of Par6 in U87MG and T98G glioma cells (n = 3 replicates). Bioluminescence images ( E ) and quantification ( F ) of xenografts derived from U87MG glioma cells in the LVCtrl and LVPar6 groups ( n = 3 per group) at 7 and 21 days after implantation. G Survival analysis of the mice in the LVCtrl and LVPar6 groups (n = 5 per group). H Immunohistochemical examination and quantification of Par6 overexpression (LVPar6) and control (LVCtrl) U87MG- xenografts ( n = 3). Bioluminescence images ( I ) and quantification ( J ) of xenografts derived from Par6-downregulated T98G cells ( n = 3 for each group). K Representative images of immunohistological staining for Par6 expression in human glioma specimens. L Quantification of Par6 expression in human glioma specimens of grades I ( n = 25), II ( n = 80), III ( n = 51) and IV ( n = 24). M Quantification of the correlation between Par6 expression and glioma recurrence. N Kaplan‒Meier overall and disease-free survival curves of human glioma samples with high and low expression of Par6. *P < 0.05, **P < 0.01, and ***P < 0.001; ns not significant. LVPar6, Par6-overexpressing group; shPar6, Par6-knockdown group; shCtrl or LVCtrl control group.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: Expressing, Over Expression, Knockdown, Derivative Assay, Immunohistochemical staining, Control, Staining

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: A GSEA enrichment plot of Par6 expression in the context of stemness maintenance in glioma cells. B The determination and quantification of Par6 expression in GSCs and the control groups from U87MG, T98G, and GBM1 cell lines. C The ELDA assay of the Par6-overexpressing, Par6-knockdown groups, and the control groups of GSCs from U87MG and T98G cells. D Representative images and quantification of the tumor sphere assay of the Par6-overexpressing or Par6-knockdown groups of U87MG-GSCs and T98G-GSCs. Scale bar, 10 μm. E Determination of the expression and quantification of stemness biomarkers, including NESTIN, CD133, OCT4 and SOX2, in Par6-overexpressing and knockdown U87MG and T98G glioma cell lines. *P < 0.05, **P < 0.01, and ***P < 0.001. LVPar6, Par6-overexpressing group; shPar6, Par6-knockdown group; shCtrl or LVCtrl, control group.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: Expressing, Control, Knockdown

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: A Determination and quantification of SOX2 protein overexpression and knockdown in U87MG and T98G glioma cell lines. B The ELDA assay of the SOX2-overexpressing, SOX2-knockdown groups, and the control groups of GSCs from U87MG and T98G cells. C Representative images and quantification of tumor sphere formation assays of U87MG and T98G glioma cell lines with SOX2 overexpression or knockdown. Scale bar, 10 μm. D Determination and quantification of the expression of NESTIN, CD133 and OCT4 in SOX2-overexpressing and SOX2-knockdown glioma cells. E ELDA assay showing the inhibition of SOX2 expression reversely regulates Par6 overexpression-enhanced self-renewal abilities in GSCs of U87MG and T98G cells. F Representative images and quantification of tumor sphere formation assay in Par6-overexpressing GSCs enriched from U87MG and T98G cells with or without transfection of SOX2 siRNAs. G Determination and quantification of the expression of NESTIN, CD133, and OCT4 in Par6-overexpressing U87MG and T98G cell lines after transfecting SOX2 siRNAs. *P < 0.05, **P < 0.01, and ***P < 0.001; ns not significant. LVPar6, Par6-overexpressing group; LVSOX2, SOX2-overexpressing group; shSOX2, SOX2-knockdown group; shCtrl or LVCtrl, control group; siSOX2, SOX2 siRNA transfecting group; siCtrl, control siRNA transfecting group.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: Over Expression, Knockdown, Control, Expressing, Inhibition, Tube Formation Assay, Transfection

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: A Representative images of immunofluorescence staining showing the colocalization of Par6 (green) and SOX2 (red) in the cytoplasm and nucleus (DAPI, blue) of glioma cells. Scale bar, 20 μm. B Representative image (upper panel) and quantitative line chart of fluorescence signal positioning analysis (lower panel) of the expression patterns of Par6 (green) and SOX2 (red) at higher magnification in the U87MG cells described in the dotted box in ( A ). C Schematic diagram showing the binding sites of Par6 and SOX2 predicted using AlphaFold Protein Structure Database. D Glioma cell lysates were subjected to coimmunoprecipitation with anti-SOX2 or anti-Par6 antibodies. The input lysate (Input) and pulldown sample (anti-SOX2 or anti-Par6) were subjected to immunoblotting. IgG was used as the negative control. E The co-immunoprecipitation between Par6 and other key factors for stemness maintenance of GSCs, including NESTIN, CD133, and OCT4. IgG was used as the negative control. F GST pull-down assay showing the direct interaction between the His-tagged SOX2 fusion protein and the GST-Par6 fusion protein. GST was utilized as the negative control.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: Immunofluorescence, Staining, Fluorescence, Expressing, Binding Assay, Western Blot, Negative Control, Immunoprecipitation, Pull Down Assay

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: A Schematic diagram of the identification of the specific peptide inhibiting the Par6/SOX2 interaction using a phage library. Three specific peptides that inhibit the Par6/SOX2 interaction (named Par6i-P1, Par6i-P2, and Par6i-P3) were identified by phage display. B Determination of cell viability after treatment with different concentrations of these three specific inhibitory peptides (0, 200, and 400 μg/ml) for 48 h ( n = 3 replicates). C In vitro evaluation of the inhibition of the Par6/SOX2 interaction by treatment with these specific inhibitory peptides ( n = 3 replicates). D Co-IP assay showing the inhibitory effect of Par6i-P1 on Par6/SOX2 interaction in the lysates from glioma cells treated with the three identified inhibitory peptides (400 μg/ml) for 48 h. The lysate (Input) was co-immunoprecipitated with anti-Par6 antibody, and the pulldown sample was subjected to immunoblotting with anti-Par6, anti-SOX2, or anti-GAPDH antibody, respectively. IgG was used as the negative control. E Binding of Par6i-P to the active site of Par6 predicted using AlphaFold Protein Structure Database. F Representative images and quantification of tumor sphere formation assays in GSCs derived from U87, T98G, and GBM1 glioma cell lines treated with or without the Par6i-P1 inhibitory peptide. Scale bar, 10 μm. G ELDA showing that Par6i-P1 treatment (400 μg/ml) on the self-renewal ability of GSCs isolated from different glioma cell lines. H The determination and quantification of the regulation of Par6, SOX2, NESTIN, CD133 and OCT4 protein expression in different glioma cells after treatment with the Par6i-P1 peptide (400 μg/ml). *P < 0.05, **P < 0.01, and ***P < 0.001; ns, not significant.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: In Vitro, Inhibition, Co-Immunoprecipitation Assay, Immunoprecipitation, Western Blot, Negative Control, Binding Assay, Derivative Assay, Isolation, Expressing

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: A Overlapping DEGs enriched in the regulated Par6 expression subgroup of glioma samples from three public datasets (TCGA-LGG + GBM, CGGA_mRNAseq_325, and CGGA_mRNAseq_693). B KEGG enrichment analysis of the overlapping DEGs described in ( A ). C Volcano plot showing the DEGs in the LVCtrl and LVPar6 groups of U87MG cells. The threshold for differential expression was set at |log 2 fold change| > 2 and P < 0.05. D KEGG analysis of the DEGs described in ( C ). E Representative images and quantification of the tumor spheres formation by Par6-overexpressing and control GSCs from glioma cells treated with or without 20 μM Capivasertib. Scale bar, 10 μm. F ELDA of the self-renewal ability of Par6-overexpressing and control GSCs derived from U87MG and T98G cells treated with or without Capivasertib. G Determination of the expression of Par6, SOX2, and PI3K/AKT signaling pathway-related factors in Par6-overexpressing and control U87MG and T98G cells. H Quantification of the activation of the PI3K/AKT signaling pathway in Par6-overexpressing U87MG and T98G cells ( n = 3 replicates). I Determination and quantification of the regulation of NESTIN, CD133, and OCT4 expression, as well as the activation of AKT signaling pathway, in Par6-overexpressing U87MG and T98G cells with or without Capivasertib treatment. *P < 0.05, **P < 0.01, and ***P < 0.001; ns not significant. LVPar6, Par6-overexpressing group; LVCtrl, control group.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: Expressing, Quantitative Proteomics, Control, Derivative Assay, Activation Assay

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: A Intersection analysis of genes associated with putative SOX2 targets from the GTRD and ChIP-Atlas databases and PI3K/AKT signaling pathway-related genes. The thresholds for screening target genes in the GTRD database were set to a site count > 2 and a CHIP-Atlas selection distance from the TSS of ± 1 kb. B The determination of mRNA levels of the potential downstream target genes of SOX2 in Par6-overexpressing and control groups of U87MG cells. C Determination and quantification of EGFR expression in Par6-overexpressing U87MG and T98G cells. D SOX2 binding motifs predicted by the JASPAR database. E Potential binding sites for the transcription factor SOX2 in the EGFR promoter predicted using the JASPAR database. F ChIP-qPCR analysis of the identified SOX2-binding site in the EGFR promoter region in U87MG cells ( n = 3 replicates). IgG was used as the negative control. G The regulation of mRNA levels of EGFR in SOX2-overexpressing and control groups of U87MG cells. ChIP-qPCR analyses of the enrichment in different SOX2-binding sites of the EGFR promoter region in Par6-overexpressing ( H ) and Par6-knockdown group ( I ) in glioma cells ( n = 3 replicates). J ChIP-qPCR analyses showing that none of SOX2-binding sites of the EGFR promoter region were detected in Par6 antibody-enriched precipitation in U87MG cells ( n = 3 replicates). The inhibition of SOX2 expression suppresses Par6-promoted EGFR expression at transcriptional ( K ) and translational ( L ) levels ( n = 3 replicates). M Determination and quantification of EGFR expression in Par6-overexpressing U87MG and T98G cells with or without Par6i-P1 treatment (400 μg/ml). N Determination and quantification of activation of AKT signaling pathways in Par6-overexpressing U87MG and T98G after Par6i-P1 treatment (400 μg/ml). O Determination and quantification of the degradation of EGFR in the Par6-overexpressing and control groups of U87MG cells stimulated with EGF at different time points (0, 2, and 4 h). P Determination and quantification of the expression of total AKT and GSK3β and their phosphorylation in Par6-overexpressing and control U87MG cells treated with EGF at different time points (0, 2, and 4 h). *P < 0.05, **P < 0.01, and ***P < 0.001. LVPar6, Par6-overexpressing group; shPar6, Par6-knockdown group; shCtrl or LVCtrl, control group; siSOX2, SOX2 siRNA transfecting group; siCtrl, control siRNA transfecting group.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: Selection, Control, Expressing, Binding Assay, ChIP-qPCR, Negative Control, Knockdown, Inhibition, Activation Assay, Protein-Protein interactions, Phospho-proteomics

Journal: Oncogene

Article Title: Par6/SOX2 interact to modulate stemness maintenance in glioma by regulating the EGFR/PI3K/AKT signaling cascade

doi: 10.1038/s41388-025-03595-7

Figure Lengend Snippet: Representative images ( A ) and quantification ( B ) of SOX2 expression in glioma specimens with low and high Par6 expression. Scale bar, 100 μm. C Kaplan‒Meier survival curves of the disease-free survival or overall survival of different subgroups (Par6-high/SOX2-low, Par6-high/SOX2-high, Par6-low/SOX2-high, and Par6-low/SOX2-low) of glioma patients. D Pearson’s correlation analysis between Par6 and SOX2 expression in the glioma tissue sample cohort. Univariate ( E ) and multivariate Cox analyses ( F ) were performed on the expression of Par6 and SOX2 and the clinicopathological parameters of glioma samples. G Schematic diagram of the molecular mechanism by which Par6 interacts with SOX2 to activate the EGFR/PI3K/AKT signaling cascade, which results in stemness maintenance and chemotherapy resistance in glioma.

Article Snippet: Par6 mRNA expression in 17 cancers in TCGA database and SOX2 protein expression in normal tissues and gliomas were downloaded from the Human Protein Atlas (HPA, https://www.proteinatlas.org/ ).

Techniques: Expressing