Journal: CytoJournal

Article Title: Mechanism of 15-hydroxyprostaglandin dehydrogenase protein inhibiting cervical cancer cell proliferation through downregulation of the notch1 signaling pathway

doi: 10.25259/Cytojournal_57_2025

Figure Lengend Snippet: 15-PGDH downregulation and activation of the Notch1 signaling pathway in human CC cells. (a) WB analysis of the expression levels of 15-PGDH and Notch1 signaling pathway-related proteins in HcerEpiC and CC cells (HeLa, Caski, ME180). (b-e) Quantitative analysis of the protein levels of 15-PGDH and Notch1 signaling pathway-related proteins. (f) RT-qPCR analysis of 15-PGDH messenger RNA levels in the cells. n = 3, ✶ P < 0.05, ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001 versus HcerEpiC. HcerEpiC: Human cervical epithelial cells, HeLa: Henrietta Lacks cells, Caski: Cervical squamous carcinoma cells, ME180: Human cervical epidermoid carcinoma cells, 15-PGDH: 15-hydroxyprostaglandin dehydrogenase, Notch1: Notch receptor 1, Jagged1: Jagged canonical Notch ligand 1, Hes1: Hes family bHLH transcription factor 1, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, CC: Cervical cancer, WB: Western blotting.

Article Snippet: The cells were divided into three groups: Control group: Cells were treated with solvent only (no intervention); SW033291 inhibitor group: Cells were treated with 0.1 mM SW033291 (HY-16968, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h; and 15-PGDH group: Cells were treated with 10 μg/mL of 15-PGDH protein (HY-P75547A, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h. Subsequently, the HeLa and Caski cells were categorized into four groups to comprehensively assess the influence of 15-PGDH protein on Notch1 overexpression-induced CC cell proliferation and Notch activation: Control group; 15-PGDH group; Notch1-OE group: Notch1 gene overexpression was induced in the HeLa and Caski cells through lentiviral-mediated gene transfection, and the cells were cultured for 48 h (the full-length Notch1 overexpression plasmid was provided by Guangzhou Ruibo Biotechnology Co., Ltd.

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: CytoJournal

Article Title: Mechanism of 15-hydroxyprostaglandin dehydrogenase protein inhibiting cervical cancer cell proliferation through downregulation of the notch1 signaling pathway

doi: 10.25259/Cytojournal_57_2025

Figure Lengend Snippet: Effect of 15-PGDH protein or its inhibitor on Notch1 signaling pathway activation in HeLa and Caski Cells. (a) WB assessment of 15-PGDH and Notch1 pathway proteins expression in HeLa cells. (b-e) Quantitative assessment of 15-PGDH and Notch1 pathway proteins amounts in HeLa cells. (f) WB assessment of 15-PGDH and Notch1 pathway proteins expression in Caski cells. (g-j) Quantitative assessment of 15-PGDH and Notch1 pathway proteins amounts in Caski cells. n = 3, ✶ P < 0.05, ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001 versus control. HeLa: Henrietta Lacks cells, Caski: Cervical squamous carcinoma cells, 15-PGDH: 15-hydroxyprostaglandin dehydrogenase, Notch1: Notch receptor 1, Jagged1: Jagged canonical Notch ligand 1, Hes1: Hes family bHLH transcription factor 1, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, WB: Western blotting.

Article Snippet: The cells were divided into three groups: Control group: Cells were treated with solvent only (no intervention); SW033291 inhibitor group: Cells were treated with 0.1 mM SW033291 (HY-16968, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h; and 15-PGDH group: Cells were treated with 10 μg/mL of 15-PGDH protein (HY-P75547A, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h. Subsequently, the HeLa and Caski cells were categorized into four groups to comprehensively assess the influence of 15-PGDH protein on Notch1 overexpression-induced CC cell proliferation and Notch activation: Control group; 15-PGDH group; Notch1-OE group: Notch1 gene overexpression was induced in the HeLa and Caski cells through lentiviral-mediated gene transfection, and the cells were cultured for 48 h (the full-length Notch1 overexpression plasmid was provided by Guangzhou Ruibo Biotechnology Co., Ltd.

Techniques: Activation Assay, Expressing, Control, Western Blot

Journal: CytoJournal

Article Title: Mechanism of 15-hydroxyprostaglandin dehydrogenase protein inhibiting cervical cancer cell proliferation through downregulation of the notch1 signaling pathway

doi: 10.25259/Cytojournal_57_2025

Figure Lengend Snippet: 15-PGDH protein inhibits Notch1 overexpression-induced proliferation of HeLa and Caski cells. (a) WB analysis of Notch1 overexpression in HeLa and Caski cells. (b) Quantitative assessment of Notch1 overexpression abundance in HeLa and Caski cells. (c and d) CCK-8 assay to detect the proliferation of HeLa and Caski cells. (e) Transwell migration assay to assess the migration ability of HeLa and Caski cells. Magnification: ×200, scale bar: 50 μm. (f and g) Statistical outcomes of the Transwell migration test. (h) EdU assay analysis of the HeLa cell proliferation rate. Magnification: ×200, scale bar: 50 μm. (i) Statistical analysis of the EdU-positive rate in HeLa cells. (j) EdU assay analysis of the Caski cell proliferation rate. Magnification: ×200, scale bar: 50 μm. (k) Statistical analysis of the EdU-positive rate in Caski cells. (l and m) Caspase activity in HeLa and Caski cells was detected using a caspase assay kit. n = 3, ✶ ✶ P < 0.01 and ✶ ✶ ✶ P < 0.001 versus Control; ### P < 0.001 versus 15-PGDH; ^^ P < 0.01 and ^^^ P < 0.001 versus Notch1-OE. HeLa: Henrietta Lacks cells, Caski: cervical squamous carcinoma cells, 15-PGDH: 15-hydroxyprostaglandin dehydrogenase, Notch1: Notch receptor 1, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, EdU: 5-ethynyl-2’-deoxyuridine, DAPI: 4',6-diamidino-2-phenylindole, Caspase-3: Cysteine-aspartic protease 3, Caspase-8: Cysteine-aspartic protease 8, and Caspase-9: Cysteine-aspartic protease 9, WB: Western blotting, CCK-8: Cell counting kit-8.

Article Snippet: The cells were divided into three groups: Control group: Cells were treated with solvent only (no intervention); SW033291 inhibitor group: Cells were treated with 0.1 mM SW033291 (HY-16968, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h; and 15-PGDH group: Cells were treated with 10 μg/mL of 15-PGDH protein (HY-P75547A, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h. Subsequently, the HeLa and Caski cells were categorized into four groups to comprehensively assess the influence of 15-PGDH protein on Notch1 overexpression-induced CC cell proliferation and Notch activation: Control group; 15-PGDH group; Notch1-OE group: Notch1 gene overexpression was induced in the HeLa and Caski cells through lentiviral-mediated gene transfection, and the cells were cultured for 48 h (the full-length Notch1 overexpression plasmid was provided by Guangzhou Ruibo Biotechnology Co., Ltd.

Techniques: Over Expression, CCK-8 Assay, Transwell Migration Assay, Migration, EdU Assay, Activity Assay, Caspase Assay, Control, Western Blot, Cell Counting

Journal: CytoJournal

Article Title: Mechanism of 15-hydroxyprostaglandin dehydrogenase protein inhibiting cervical cancer cell proliferation through downregulation of the notch1 signaling pathway

doi: 10.25259/Cytojournal_57_2025

Figure Lengend Snippet: 15-PGDH protein inhibits Notch activation induced by Notch1 overexpression in HeLa and Caski cells. (a) WB analysis of 15-PGDH and proteins associated with the Notch1 pathway in HeLa cells. (b-e) Quantitative assessment of 15-PGDH and Notch1 pathway-related proteins expression levels. (f) WB analysis of 15-PGDH and proteins associated with the Notch1 pathway in Caski cells. (g-j) Quantitative assessment of 15-PGDH and Notch1 pathway-related protein expression levels. n = 3, ✶ ✶ ✶ P < 0.001 versus Control; ### P < 0.001 versus 15-PGDH; ^^^ P < 0.001 versus Notch1-OE. HeLa: Henrietta Lacks Cells, Caski: Cervical squamous carcinoma cells, 15-PGDH: 15-hydroxyprostaglandin dehydrogenase, Notch1: Notch receptor 1, Jagged1: Jagged canonical Notch ligand 1, Hes1: Hes family bHLH transcription factor 1, GAPDH: Glyceraldehyde-3-phosphate dehydrogenase, WB: Western blotting.

Article Snippet: The cells were divided into three groups: Control group: Cells were treated with solvent only (no intervention); SW033291 inhibitor group: Cells were treated with 0.1 mM SW033291 (HY-16968, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h; and 15-PGDH group: Cells were treated with 10 μg/mL of 15-PGDH protein (HY-P75547A, MedChem Express, Monmouth Junction, NJ, USA) and cultured for 24 h. Subsequently, the HeLa and Caski cells were categorized into four groups to comprehensively assess the influence of 15-PGDH protein on Notch1 overexpression-induced CC cell proliferation and Notch activation: Control group; 15-PGDH group; Notch1-OE group: Notch1 gene overexpression was induced in the HeLa and Caski cells through lentiviral-mediated gene transfection, and the cells were cultured for 48 h (the full-length Notch1 overexpression plasmid was provided by Guangzhou Ruibo Biotechnology Co., Ltd.

Techniques: Activation Assay, Over Expression, Expressing, Control, Western Blot

Journal: Molecular Medicine Reports

Article Title: GPM6B inhibits tumor progression by targeting HPGD in lung adenocarcinoma

doi: 10.3892/mmr.2025.13618

Figure Lengend Snippet: GPM6B regulates HPGD expression. (A) DEGs identified via transcriptome sequencing following GPM6B overexpression. (B) Reverse transcription-quantitative PCR validation of DEGs. (C) Western blot analysis of HPGD expression. (D) OS plot of HPGD in LUAD. (E) Nomogram was used to predict the OS probability of patients with LUAD at 1, 3 and 5 years. mRNA expression levels of HPGD in LUAD and paired adjacent normal tissue in (F) The Cancer Genome Atlas and (G) GSE10072 and GSE115002 datasets. (H) IHC analysis of HPGD in LUAD and adjacent normal tissues. Scale bar, 100 µm. *P<0.05 vs. Normal; **P<0.01, ***P<0.01, ****P<0.0001 vs. Control. (G, H) HPGD, 15-hydroxy prostaglandin dehydrogenase; DEG, differentially expressed gene; OS, overall survival; LUAD, lung adenocarcinoma; IHC, immunohistochemistry; ns, not significant.

Article Snippet: The clinical samples were incubated with antibodies against GPM6B (cat. no. PC11847S; Abmart Pharmaceutical Technology Co., Ltd.; 1:400) and HPGD (cat. no. 66798-1-Ig; Proteintech Group, Inc.; 1:400), while the mouse tissue samples were incubated with Ki67 primary antibody (cat. no. A11390; ABclonal Biotech Co., Ltd.; 1:500) overnight at 4°C.

Techniques: Expressing, Sequencing, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Biomarker Discovery, Western Blot, Control, Immunohistochemistry

Journal: Molecular Medicine Reports

Article Title: GPM6B inhibits tumor progression by targeting HPGD in lung adenocarcinoma

doi: 10.3892/mmr.2025.13618

Figure Lengend Snippet: Association between GPM6B and HPGD. (A) Western blot showing the expression of GPM6B and HPGD in 293T cells transfected with HA-tagged GPM6B plasmid. Association between GPM6B and HPGD expression in (B) TCGA and (C) GSE10072 and GSE115002 datasets. HPGD, 15-hydroxy prostaglandin dehydrogenase; TGCA, The Cancer Genome Atlas; GPM6B, Glycoprotein M6B; IP, Immunoprecipitation; TPM, Transcripts Per Million.

Article Snippet: The clinical samples were incubated with antibodies against GPM6B (cat. no. PC11847S; Abmart Pharmaceutical Technology Co., Ltd.; 1:400) and HPGD (cat. no. 66798-1-Ig; Proteintech Group, Inc.; 1:400), while the mouse tissue samples were incubated with Ki67 primary antibody (cat. no. A11390; ABclonal Biotech Co., Ltd.; 1:500) overnight at 4°C.

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Molecular Medicine Reports

Article Title: GPM6B inhibits tumor progression by targeting HPGD in lung adenocarcinoma

doi: 10.3892/mmr.2025.13618

Figure Lengend Snippet: GPM6B overexpression activates p53 signaling pathway. (A) GO, (B) KEGG and (C) Gene Set Enrichment Analysis. (D) Western blot images revealed changes in the expression of GPM6B, HPGD, P53, CCND1, Bcl2 and Bax. ****P<0.0001 vs. Control. (D) GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; GPM6B, Glycoprotein M6B; HPGD, 15-hydroxy prostaglandin dehydrogenase; CCND1, cyclin D1; NES, normalized enrichment score.

Article Snippet: The clinical samples were incubated with antibodies against GPM6B (cat. no. PC11847S; Abmart Pharmaceutical Technology Co., Ltd.; 1:400) and HPGD (cat. no. 66798-1-Ig; Proteintech Group, Inc.; 1:400), while the mouse tissue samples were incubated with Ki67 primary antibody (cat. no. A11390; ABclonal Biotech Co., Ltd.; 1:500) overnight at 4°C.

Techniques: Over Expression, Western Blot, Expressing, Control

Journal: Nature Communications

Article Title: Modulating the PPARγ pathway upregulates NECTIN4 and enhances chimeric antigen receptor (CAR) T cell therapy in bladder cancer

doi: 10.1038/s41467-025-62710-0

Figure Lengend Snippet: a NECTIN4 mRNA levels in RT112 cells treated with 1μM rosiglitazone and T0070907 (T007) for 72 h. Data are presented as mean ± SEM, n = 6 for DMSO and rosiglitazone, n = 4 for T0070907, all biological replicates. b Western blot for NECTIN4 and HPGD in RT112 cells treated with T0070907, rosiglitazone, and pioglitazone at indicated concentrations for 72 h. This was repeated n = 3 independent times with similar results. c, d NECTIN4 surface staining ( c ) and quantification of median fluorescence intensify (MFI) ( d ) in RT112 cells treated with 1μM rosiglitazone and T0070907 for 72 h. Data are presented as mean ± SEM, n = 4 for DMSO and T0070907, n = 5 for rosiglitazone, all biological replicates. e Western blot for NECTIN4 and HPGD in RT112 cells treated with rosiglitazone for 72 h across a dose series (starting at 1 μM on the right most lane with serial 2-fold dilutions to the left). This was repeated n = 3 independent times with similar results. f Dose response curves of total NECTIN4 and HPGD protein expression after 72 h rosiglitazone treatment in RT112 cells. Data are presented as mean values. g NECTIN4 surface staining in RT112 cells treated with rosiglitazone for 72 h across a dose series. Dose series starts at 1μM with serial 2-fold dilutions. h Dose response curves of surface NECTIN4 and TROP2 expression for rosiglitazone in RT112 cells after 72 h. Data are presented as mean ± SEM, n = 2 biological replicates. i Western blots for NECTIN4, PPARγ and FABP4 in RT112 cells expressing sgRNAs against GAL4 (control), NECTIN4 , and two unique PPARG guides. This was repeated n = 3 independent times with similar results. j Schematic of select predicted PPARG binding sites (labeled 1 and 2) on the NECTIN4 promoter region (500 to −2k) by FIMO using transcription binding motif matrices JASPAR 2018. Created with BioRender. Chang (2025) https://BioRender.com/k3j6fgi . k Chromatin from RT112 cells was precipitated using antibodies against PPARG or IgG. Primers targeting putative PPARG binding motifs 1 and 2 from (j) were used for PCR and the PCR product was visualized by gel electrophoresis. l Quantitative analysis of ChIP-qPCR experiments. Results are represented as fold-enrichment relative to IgG control. Data are presented as mean ± SEM, n = 3 biological replicates. For panels ( a) and ( d ), ordinary one-way ANOVA with Sidak’s multiple comparison was used. For panel ( l ), two-sided, unpaired Student’s t test was used for each primer set. Vinculin shown as a loading control for panels ( b ) and ( e ) and GAPDH for panel ( i ). Source data are provided as a Source Data file.

Article Snippet: Antibodies against GAPDH (Cell Signaling Technology, #2118, clone 14C10, 1:4000), Vinculin (Cell Signaling Technology, #13901, clone E1E9V, 1:2000), NECTIN4 (abcam #192033, clone EPR14613-68, 1:1000), PPARγ (Cell Signaling Technology, #2435, clone C26H12, 1:1000), HPGD (R&D Systems, AF5660, 1:1000), FABP4 (Cell Signaling Technology, #2120, 1:1000), HRP-anti-rabbit (Cell Signaling Technology, #7074, 1:8000) and HRP-anti-goat (abcam, #ab6741, 1:8000) were used according to the manufacturer’s recommended dilutions.

Techniques: Western Blot, Staining, Fluorescence, Expressing, Control, Binding Assay, Labeling, Nucleic Acid Electrophoresis, ChIP-qPCR, Comparison

Journal: eLife

Article Title: Oligodendrocyte-lineage cell exocytosis and L-type prostaglandin D synthase promote oligodendrocyte development and myelination

doi: 10.7554/eLife.77441

Figure Lengend Snippet: ( A–D ) Differentiated oligodendrocytes, OPCs, astrocytes, and microglia were purified by immunopanning from whole brains of P17 PD:ibot and littermate control mice. Anti-GalC hybridoma was used to purify differentiated oligodendrocytes; anti-O4 hybridoma was used to isolate OPCs; anti-HepaCAM antibody was used to purify astrocytes; anti-CD45 antibody was used to isolate microglia. More details can be found in the method section. Gene expression was determined by RNA-seq. Genes exhibiting significant changes (P-value adjusted for multiple comparisons, Padj <0.05, fold change >1.5) are shown in red. ( E–H ) Examples of mature oligodendrocyte marker gene expression by oligodendrocytes purified from PD:ibot and control mice at P17. N=3 mice per group. Significance is determined by DESeq2. Plp1 : 1586±375.9 in control and 729.3±152.4 in PD:ibot, p=3.96 × 10 –13 ; Mbp : 135.2±12.8 in control and 89.8±13.5 in PD:ibot, p=4.76 × 10 –5 ; Aspa : 5.9±1.7 in control and 2.0±0.6 in PD:ibot, p=0.003; Mobp : 274.4±40.1 in control and 97.5±24.0 in PD:ibot, p=0.0004. ( I–L ) Examples of GO terms associated with genes upregulated in oligodendrocytes ( I ), downregulated in oligodendrocytes ( J ), upregulated in OPCs ( K ), and downregulated in microglia ( L ). There are no GO terms significantly associated with genes downregulated in OPCs or upregulated in microglia. ( M, N ) Expression of Ptgds by OPCs and oligodendrocytes purified from PD:ibot and control mice at P17. N=3 mice per group. Significance is determined by DESeq2. Ptgds in OPCs: 11.6±4.3 in control and 34.5±10.5 in PD:ibot, p=1.79 × 10 –12 ; Ptgds in oligodendrocytes: 4.1±0.3 in control and 39.1±13.9 in PD:ibot, p=3.14 × 10 –8 . ( O ) Expression of Hpgd by microglia purified from PD:ibot and control mice at P17. ( E–H, M–O ) Expression is shown in RPKM. N=3 mice per group. Significance is determined by DESeq2. Hpgd in microglia: 11.7±0.9 in control and 6.1±0.7 in PD:ibot, p=6.34 × 10 –19 . Figure 4—source data 1. It contains original data points for .

Article Snippet: HPGD protein (R&D system, cat#5660-DH-010) was added to the oligodendrocyte culture medium at 3 μM and 6 μM daily.

Techniques: Purification, Control, Gene Expression, RNA Sequencing, Marker, Expressing

Journal: eLife

Article Title: Oligodendrocyte-lineage cell exocytosis and L-type prostaglandin D synthase promote oligodendrocyte development and myelination

doi: 10.7554/eLife.77441

Figure Lengend Snippet: ( A ) Immunoblot of secreted and intracellular L-PGDS protein from oligodendrocyte cultures from PD:ibot and littermate control mice. ( B ) Quantification of the immunoblot signal intensity of secreted L-PGDS. N=10 mice per group. Paired two-tailed T-test. Secreted L-PGDS intensity: 3.0±0.4 in control and 2.2±0.4 in PD:ibot, p=0.0104. ( C ) Quantification of the ratio of secreted and intracellular L-PGDS. N=10 mice per group. Paired two-tailed T-test. Ratio of secreted and intracellular L-PGDS: 37.2±10.4 in control and 18.9±3.7 in PD:ibot, p=0.05. ( D ) Oligodendrocyte cultures from wild-type mice after 7 days of differentiation in the presence and absence of the L-PGDS inhibitor AT-56. Red: MBP immunofluorescence. Blue: CellMask, which labels all cells. Scale bars: 50 μm. ( E ) Quantification of cells with lamellar morphology. One-way ANOVA with Benjamini, Krieger, and Yekutieli’s two-stage linear step-up FDR procedure for multiple comparisons. N=3 cultures from 3 mice per group. Lamellar cells%: DMSO control: 28±0.8; 1 μM AT-56: 15±1.6, p=0.024; 5 μM AT-56: 7.3±1.3, p=0.0068. ( F ) Oligodendrocyte cultures from wild-type mice after 7 days of differentiation in the presence and absence of HPGD. Green: Membrane version of CellMask, which labels the membranes of all cells. Blue: DAPI. Scale bars: 50 μm. ( G ) Quantification of cells with lamellar morphology. One-way ANOVA with Benjamini, Krieger, and Yekutieli’s two-stage linear step-up FDR procedure for multiple comparisons. N=7 cultures from 6 mice per group. Lamellar cells%: 0 μM HPGD: 59.10±5.33; 3 μM HPGD: 50.09±5.80, p=0.034; 6 μM HPGD: 42.20±5.87, p=0.0090. Figure 7—source data 1. It contains original data points for . Figure 7—source data 2. It contains figures with the uncropped blots with the relevant bands clearly labelled ( A ) and original blot images ( B and C ). ( A ) Labelled original data for . ( B ) Original blot for L-PGDS in . ( C ) Original blot for GAPDH in .

Article Snippet: HPGD protein (R&D system, cat#5660-DH-010) was added to the oligodendrocyte culture medium at 3 μM and 6 μM daily.

Techniques: Western Blot, Control, Two Tailed Test, Immunofluorescence, Membrane

Journal: eLife

Article Title: Oligodendrocyte-lineage cell exocytosis and L-type prostaglandin D synthase promote oligodendrocyte development and myelination

doi: 10.7554/eLife.77441

Figure Lengend Snippet:

Article Snippet: HPGD protein (R&D system, cat#5660-DH-010) was added to the oligodendrocyte culture medium at 3 μM and 6 μM daily.

Techniques: Recombinant, Purification, Software, Sequencing, Staining, Membrane