Journal: bioRxiv

Article Title: Rewiring of master transcription factor cistromes during high-grade serous ovarian cancer development

doi: 10.1101/2023.04.11.536378

Figure Lengend Snippet: (A) Unsupervised hierarchical clustering of MECOM , PAX8 , SOX17 and WT1 mRNA expression in the pan-normal GTEx dataset. TCGA data was clustered based on the 5 main clusters of TF expression from GTEx. MTF low = Cluster of tissues that did not or lowly expressed MTFs, MTF high = cluster of tissues that moderately or highly expressed all four MTFs, M high = Cluster of tissues that highly expressed MECOM, M&P high = cluster of tissues that highly expressed both MECOM and PAX8, P high = cluster of tissues that expresses PAX8 and S high = cluster of tissues that expresses SOX17). (B) A boxplot of the average Pearson correlation values of MECOM, PAX8, SOX17 and WT1. Ranked from highest average Pearson correlation value to lowest. (C) A boxplot representing the mean positivity rate of MECOM, PAX8, SOX17 and WT1 expression in each histotype. In B and C, the limits of the boxes represent the interquartile range, and the limits of error bars represent the minimum and maximum value without outliers (D) Ratio of samples with number of co-stained TFs based on a 0.1 positivity rate threshold.

Article Snippet: Isolated nuclei from FT282 and KURAMOCHI were bound to Concalavin A beads and incubated overnight at 4°C with primary antibody for H3K27me3 (Cell Signaling, RRID:AB_2798370), H3K27ac (Diagenode, RRID:AB_2637079), MECOM (Cell Signaling, AB_2184098), PAX8 (Novus, RRID:AB_2283498), SOX17 (Abcam, RRID:AB_2801385), WT1 (Santa Cruz, RRID:AB_632611).

Techniques: Expressing, Staining

Journal: bioRxiv

Article Title: Rewiring of master transcription factor cistromes during high-grade serous ovarian cancer development

doi: 10.1101/2023.04.11.536378

Figure Lengend Snippet: Landscapes of active chromatin and chromatin loops (based on Hi-C maps) in FTSECs and HGSCs. (A) MECOM, (B) PAX8, (C) SOX17 and (D) WT1 .

Article Snippet: Isolated nuclei from FT282 and KURAMOCHI were bound to Concalavin A beads and incubated overnight at 4°C with primary antibody for H3K27me3 (Cell Signaling, RRID:AB_2798370), H3K27ac (Diagenode, RRID:AB_2637079), MECOM (Cell Signaling, AB_2184098), PAX8 (Novus, RRID:AB_2283498), SOX17 (Abcam, RRID:AB_2801385), WT1 (Santa Cruz, RRID:AB_632611).

Techniques: Hi-C

Journal: bioRxiv

Article Title: Rewiring of master transcription factor cistromes during high-grade serous ovarian cancer development

doi: 10.1101/2023.04.11.536378

Figure Lengend Snippet: (A) TF knock-down followed by colony formation assays stained with crystal violet. Representative wells are shown. (B) Barplots representing the quantification of crystal violet staining in . Error bars represent biological replicates. (C) Dose response curves for FT246, FT282, KURAMOCHI and OVCAR4 cells treated with THZ1, THZ531 and JQ1. Error bars represent standard deviation of mean cell survival values from biological replicates. (D) RT-qPCR quantification of MECOM , PAX8 , SOX17 and WT1 expression upon THZ1 and THZ531 treatment in FT282 and OVCAR4 cells. Data for MECOM , PAX8 and SOX17 expression upon THZ1 and THZ531 treatment of OVCAR4 cells are reproduced from .

Article Snippet: Isolated nuclei from FT282 and KURAMOCHI were bound to Concalavin A beads and incubated overnight at 4°C with primary antibody for H3K27me3 (Cell Signaling, RRID:AB_2798370), H3K27ac (Diagenode, RRID:AB_2637079), MECOM (Cell Signaling, AB_2184098), PAX8 (Novus, RRID:AB_2283498), SOX17 (Abcam, RRID:AB_2801385), WT1 (Santa Cruz, RRID:AB_632611).

Techniques: Knockdown, Staining, Standard Deviation, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Rewiring of master transcription factor cistromes during high-grade serous ovarian cancer development

doi: 10.1101/2023.04.11.536378

Figure Lengend Snippet: (A) MECOM, PAX8, SOX17 and WT1 co-occupies its own and others genomic loci. ( B) MECOM, PAX8, SOX17 and WT1 co-occupy active enhancer regions across the genome. CPM-normalized CUT&RUN reads were centered on 3 kilobase windows of FTSEC or HGSC-specific PAX8 peaks. Rows are the same across feature. (C) Metagene plot, MECOM, PAX8, SOX17, WT1, H3K27ac and H3K27me3 signal centered on FT of HGSC-specific PAX8 peaks. (D) Set analysis of CUT&RUN peaks from representative MECOM, PAX8, SOX17 and WT1 samples in FT282 and KURAMOCHI. (E) Chromatin state of TF peaks categorized by number of TF overlaps. (F) MECOM, PAX8, SOX17 and WT1 co-regulation based on TF knock-down followed by RNA-seq and differential expression analysis with DESEQ2. (G) Node and edge plot representing co-regulation of each TF based on gene expression measured by RNA-seq after TF knock-down.

Article Snippet: Isolated nuclei from FT282 and KURAMOCHI were bound to Concalavin A beads and incubated overnight at 4°C with primary antibody for H3K27me3 (Cell Signaling, RRID:AB_2798370), H3K27ac (Diagenode, RRID:AB_2637079), MECOM (Cell Signaling, AB_2184098), PAX8 (Novus, RRID:AB_2283498), SOX17 (Abcam, RRID:AB_2801385), WT1 (Santa Cruz, RRID:AB_632611).

Techniques: Knockdown, RNA Sequencing, Quantitative Proteomics, Gene Expression

Journal: bioRxiv

Article Title: Rewiring of master transcription factor cistromes during high-grade serous ovarian cancer development

doi: 10.1101/2023.04.11.536378

Figure Lengend Snippet: (A) Set analysis of TF binding sites that were common or specific to FT282 or KURAMOCHI. (B) MECOM, PAX8, SOX17 and WT1 co-occupies regions that were bound in a common or context-specific manner. CPM-normalized CUT&RUN reads were centered on 3 kilobase windows of PAX8 peak start and stop positions. Rows are the same across samples. (C) Ratio of chromatin states associated with TF binding regions categorized by cellular context. (D) Ratio of FT282 and KURAMOCHI specific enhancers bound by one, two, three or four TFs. (E) BHLHE41 and PBX1 loci displaying the co-localization of MECOM, PAX8, SOX17 and WT1 at a KURAMOCHI specific super-enhancer.

Article Snippet: Isolated nuclei from FT282 and KURAMOCHI were bound to Concalavin A beads and incubated overnight at 4°C with primary antibody for H3K27me3 (Cell Signaling, RRID:AB_2798370), H3K27ac (Diagenode, RRID:AB_2637079), MECOM (Cell Signaling, AB_2184098), PAX8 (Novus, RRID:AB_2283498), SOX17 (Abcam, RRID:AB_2801385), WT1 (Santa Cruz, RRID:AB_632611).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Rewiring of master transcription factor cistromes during high-grade serous ovarian cancer development

doi: 10.1101/2023.04.11.536378

Figure Lengend Snippet: (A) Log 2 fold-change of 28,158 genes following normalization informed by ERCC spike-in RNA content, (B) Number of differentially expressed genes for each TF knock-down based on absolute log 2 fold-change ≥ 0.5. (C) Schematic to integrate differential expression, TF binding and topologically associated domain (TAD) maps. (D) Alluvial plot displaying the status of high confidence differentially expressed genes following TF depletion in FTSECs and HGSCs. (E) Heatmap and pathway analysis of genes displayed in D. (F) BRCA1 locus highlighting H3K27ac signal and TF binding at the BRCA1 promoter. (G) Log 2 fold change of BRCA1 expression following MECOM, PAX8, SOX17 and WT1 knock-down relative to scrambled controls. (H) Chromatin landscape of RUNX3 locus in FTSECs and HGSCs. (I) Log 2 fold change of RUNX3 expression following MECOM, PAX8, SOX17 and WT1 knock-down relative to scrambled controls (RNA-seq).

Article Snippet: Isolated nuclei from FT282 and KURAMOCHI were bound to Concalavin A beads and incubated overnight at 4°C with primary antibody for H3K27me3 (Cell Signaling, RRID:AB_2798370), H3K27ac (Diagenode, RRID:AB_2637079), MECOM (Cell Signaling, AB_2184098), PAX8 (Novus, RRID:AB_2283498), SOX17 (Abcam, RRID:AB_2801385), WT1 (Santa Cruz, RRID:AB_632611).

Techniques: Knockdown, Quantitative Proteomics, Binding Assay, Expressing, RNA Sequencing

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: C18-3OH modulates ABCA1 expression and cholesterol efflux in foam cells via PAX-8 upregulation. (A) RT-qPCR analysis of PAX-8 mRNA expression in foam cells treated with C18-3OH. (B) Western blot analysis of PAX-8 protein expression in foam cells treated with C18-3OH. ** P < 0.01 vs. control, n = 3. (C) Western blot detection of PAX-8 and ABCA1 protein expression in foam cells following PAX-8 knockdown. (D) Quantitative analysis of PAX-8 protein expression (Panel C). (E) RT-qPCR analysis of ABCA1 mRNA expression in foam cells after PAX-8 knockdown. (F) Quantitative analysis of ABCA1 protein expression (Panel C). (G) Effects of C18-3OH and siPAX-8 on cholesterol efflux in foam cells. * P < 0.05, ** P < 0.01 vs. siNC + DMSO; ## P < 0.01 vs. siNC + C18-3OH, n = 3. (H) NBD-cholesterol fluorescence intensity in foam cells; scale bar = 150 μm.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Knockdown, Fluorescence

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: PAX-8 binds to and regulates ABCA1 expression. (A) Predicted binding sites between PAX-8 and ABCA1 in humans. (B) Predicted binding sites between PAX-8 and ABCA1 in mice. (C) Western blot analysis of PAX-8 and ABCA1 protein expression in foam cells following PAX-8 overexpression. (D) Quantitative analysis of PAX-8 protein expression (Panel C). (E) RT-qPCR analysis of ABCA1 mRNA expression in foam cells after PAX-8 overexpression. (F) Quantitative analysis of ABCA1 protein expression (Panel C). ** P < 0.01 vs. LV-NC, n = 3. (G) ChIP-Seq identification of PAX-8 binding to the ABCA1 locus. Genomic tracks showing PAX-8 binding to the ABCA1 locus in the Input (negative control, top) and ChIP (PAX-8-enriched, bottom) groups. The y-axis indicates relative enrichment of ChIP-Seq signals. The x-axis represents genomic coordinates. (H) ChIP-qPCR validation of PAX-8 targeting regulation of ABCA1. ** P < 0.01 vs. Input, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Binding Assay, Western Blot, Over Expression, Quantitative RT-PCR, ChIP-sequencing, Negative Control, ChIP-qPCR, Biomarker Discovery

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: Effects of C18-3OH on m 6 A modification of PAX-8 mRNA in foam cells. (A) Predicted score distribution of the human PAX-8 gene query sequence. (B) Predicted score distribution of the murine PAX-8 gene query sequence. (C) Partitioned statistical plot of predicted scores for the human PAX-8 gene query sequence. (D) Partitioned statistical plot of predicted scores for the murine PAX-8 gene query sequence. (E) MeRIP-qPCR analysis of the effect of C18-3OH on m 6 A modification of PAX-8 mRNA in foam cells. ** P < 0.01 vs. control, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Modification, Sequencing, Control

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: Effects of C18-3OH on ALKBH5 expression in foam cells. (A) Bioinformatic analysis of methylation-associated proteins regulating PAX-8 in THP-1 cells. (B) RT-qPCR analysis of ALKBH5 mRNA expression in foam cells treated with C18-3OH. (C) Western blot analysis of ALKBH5 protein expression in foam cells treated with C18-3OH. (D) RT-qPCR analysis of ELAVL1 mRNA expression in foam cells treated with C18-3OH. (E) Western blot analysis of ELAVL1 protein expression in foam cells treated with C18-3OH. * P < 0.05, ** P < 0.01 vs. control, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Methylation, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: ALKBH5 mediates C18-3OH regulation of PAX-8 and ABCA1 expression, PAX-8 mRNA m 6 A modification, and cholesterol efflux. (A) Western blot analysis of ALKBH5 expression in foam cells following lentiviral transfection for ALKBH5 overexpression. ** P < 0.01 vs. control, n = 3. (B) Western blot analysis of PAX-8 and ABCA1 protein expression in foam cells following ALKBH5 overexpression. (C) RT-qPCR analysis of ABCA1 mRNA expression in foam cells after ALKBH5 overexpression. (D) Quantitative analysis of ABCA1 protein expression (Panel B). (E) RT-qPCR analysis of PAX-8 mRNA expression in foam cells after ALKBH5 overexpression. (F) Quantitative analysis of PAX-8 protein expression (Panel B). (G) MeRIP-qPCR analysis of PAX-8 mRNA m6A modification in foam cells following ALKBH5 overexpression. (H) Effect of C18-3OH and LV-ALKBH5 on cholesterol efflux in foam cells. (I) NBD-cholesterol fluorescence intensity in foam cells; scale bar = 150 μm. * P < 0.05, ** P < 0.01 vs. LV-NC + DMSO; ## P < 0.01 vs. LV-NC + C18-3OH, n = 3.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Modification, Western Blot, Transfection, Over Expression, Control, Quantitative RT-PCR, Fluorescence

Journal: Frontiers in Immunology

Article Title: 3-Hydroxystearic acid promotes cholesterol efflux and attenuates atherosclerosis via the ALKBH5/PAX-8/ABCA1 pathway

doi: 10.3389/fimmu.2026.1750021

Figure Lengend Snippet: Effects of C18-3OH on ABCA1, PAX-8, ALKBH5 expression, and PAX-8 mRNA m 6 A methylation in Aortas of apoE −/− Mice. (A) Western blot analysis of ABCA1, PAX-8, and ALKBH5 protein expression in aortas of HFD-fed apoE −/− mice treated with C18-3OH and/or ALKBH5 overexpression. (B) RT-qPCR analysis of ABCA1 mRNA expression in aortas of apoE −/− mice. (C) RT-qPCR analysis of PAX-8 mRNA expression in aortas of apoE −/− mice. (D) RT-qPCR analysis of ALKBH5 mRNA expression in aortas of apoE −/− mice. (E) Quantitative analysis of ABCA1 protein expression (Panel A). (F) Quantitative analysis of PAX-8 protein expression (Panel A). (G) Quantitative analysis of ALKBH5 protein expression (Panel A). (H) MeRIP-qPCR analysis of PAX-8 mRNA m 6 A methylation levels in aortas of apoE −/− mice across experimental groups. ** P < 0.01 vs. Control; # P < 0.05, ## P < 0.01 vs. C18-3OH + AAV-NC. n=6.

Article Snippet: Primary antibodies diluted in antibody dilution buffer were applied as follows: ABCA1 (Abcam, 1:1000), PAX-8 (Proteintech, 1:3000), ALKBH5 (Proteintech, 1:5000), ELAVL1 (Proteintech, 1:5000); GAPDH (Proteintech, 1:20, 000) and α-Tubulin (Proteintech, 1:20, 000) served as loading controls.

Techniques: Expressing, Methylation, Western Blot, Over Expression, Quantitative RT-PCR, Control

Journal: BMC Developmental Biology

Article Title: The nephrogenic potential of the transcription factors osr1, osr2, hnf1b, lhx1 and pax8 assessed in Xenopus animal caps

doi: 10.1186/1471-213X-11-5

Figure Lengend Snippet: Expression of human LHX1, PAX8 and HNF1B in animal caps . Animal caps of LHX1 and/or PAX8 (250 pg alone or 125 pg each) or HNF1B (150 pg) injected embryos were cultured in Steinberg's solution for three hours and analysed after four days by whole-mount immunostaining using a mixture of the antibodies 3G8 and 4A6. Activin A (ActA, 10 ng/ml) and retinoic acid (RA, 10 -4 M) were added as given. (A) and (B) The pronephric tissue induction in animal caps treated as indicated on the left was scored using the categories given in Figure 4A. The number of animal caps (N) is given.

Article Snippet: The full-length open reading frames of the human PAX8 (RC200651) and LHX1 (RC210977) in pCMV6 were obtained from OriGene Technologies.

Techniques: Expressing, Injection, Cell Culture, Immunostaining

Journal: BMC Developmental Biology

Article Title: The nephrogenic potential of the transcription factors osr1, osr2, hnf1b, lhx1 and pax8 assessed in Xenopus animal caps

doi: 10.1186/1471-213X-11-5

Figure Lengend Snippet: Activation of genes by HNF1B in animal caps

Article Snippet: The full-length open reading frames of the human PAX8 (RC200651) and LHX1 (RC210977) in pCMV6 were obtained from OriGene Technologies.

Techniques: Activation Assay

Journal: BMC Developmental Biology

Article Title: The nephrogenic potential of the transcription factors osr1, osr2, hnf1b, lhx1 and pax8 assessed in Xenopus animal caps

doi: 10.1186/1471-213X-11-5

Figure Lengend Snippet: Induction of the early nephrogenic transcription factors and other genes in animal caps . osr1, osr2, hnf1b and lhx1 are induced already after 1.5 hours treatment with activin A (ActA) and retinoic acid (RA) suggesting that they are direct targets, whereas pax8 is first expressed after thirteen hours implying an indirect activation. osr2 and lhx1 (highlighted in grey) are induced after three hours by activin A alone. HNF1B overexpressed in animal caps induces after seven hours osr1, osr2, hnf1a, hnf4a, wnt11b, gdnf and tfe3. After fourteen hours pax2 and esd are increased and rbms1, fgfr4c and gsc are decreased suggesting secondary effects. HNF1B induces the expression of lhx1 by an HNF1 binding site in its promoter region (arrow). Furthermore, the lhx1 target genes cer1 and chrd are induced in HNF1B injected animal caps possibly via lhx1.

Article Snippet: The full-length open reading frames of the human PAX8 (RC200651) and LHX1 (RC210977) in pCMV6 were obtained from OriGene Technologies.

Techniques: Activation Assay, Expressing, Binding Assay, Injection

Journal: Heliyon

Article Title: Prognostic value of PAX8 in small cell lung cancer

doi: 10.1016/j.heliyon.2024.e28251

Figure Lengend Snippet: Expression of PAX8 in clinicopathological characteristics of SCLC.

Article Snippet: The samples were incubated with anti-Mouse PAX8 monoclonal antibody (1:1 dilution; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd; clone number: OTI6H8) or anti-Mouse Ki-67 monoclonal antibody (1:2500 dilution; Beijing Zhongshan Golden Bridge Biotechnology; clone number: UMAB107).

Techniques: Expressing

Journal: Heliyon

Article Title: Prognostic value of PAX8 in small cell lung cancer

doi: 10.1016/j.heliyon.2024.e28251

Figure Lengend Snippet: Immunohistochemical expression of PAX8 in SCLC. (A) Extensive-stage SCLC PAX8 positive case. (B) Limited-stage small cell lung cancer PAX8 positive case. (C) Extensive-stage small cell lung cancer PAX8 negative case. (D) limited-stage small cell lung cancer PAX8 negative case.

Article Snippet: The samples were incubated with anti-Mouse PAX8 monoclonal antibody (1:1 dilution; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd; clone number: OTI6H8) or anti-Mouse Ki-67 monoclonal antibody (1:2500 dilution; Beijing Zhongshan Golden Bridge Biotechnology; clone number: UMAB107).

Techniques: Immunohistochemical staining, Expressing

Journal: Heliyon

Article Title: Prognostic value of PAX8 in small cell lung cancer

doi: 10.1016/j.heliyon.2024.e28251

Figure Lengend Snippet: Overexpression of PAX8 in HEK-293T. (A) Real-time PCR analysis of PAX8 in HEK-293T. (B) Immunohistochemical expression of PAX8 in HEK-293T.

Article Snippet: The samples were incubated with anti-Mouse PAX8 monoclonal antibody (1:1 dilution; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd; clone number: OTI6H8) or anti-Mouse Ki-67 monoclonal antibody (1:2500 dilution; Beijing Zhongshan Golden Bridge Biotechnology; clone number: UMAB107).

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Expressing

Journal: Heliyon

Article Title: Prognostic value of PAX8 in small cell lung cancer

doi: 10.1016/j.heliyon.2024.e28251

Figure Lengend Snippet: Comparative survival analyses in SCLC patients. (A) Comparison of OS between PAX8 positive and PAX8 negative group. (B) Comparison of OS between Ki-67 high and Ki-67 low group. (C) Comparison of OS between Limited-stage and Extensive-stage patients. (D) Comparison of OS among PAX8 expression and stage status combination groups. The difference in OS between groups was compared by Log-rank test, with death as the outcome.

Article Snippet: The samples were incubated with anti-Mouse PAX8 monoclonal antibody (1:1 dilution; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd; clone number: OTI6H8) or anti-Mouse Ki-67 monoclonal antibody (1:2500 dilution; Beijing Zhongshan Golden Bridge Biotechnology; clone number: UMAB107).

Techniques: Comparison, Expressing

Journal: Heliyon

Article Title: Prognostic value of PAX8 in small cell lung cancer

doi: 10.1016/j.heliyon.2024.e28251

Figure Lengend Snippet: The HR value of Ki-67, PAX8 and stage status for overall survival.

Article Snippet: The samples were incubated with anti-Mouse PAX8 monoclonal antibody (1:1 dilution; Beijing Zhongshan Golden Bridge Biotechnology Co., Ltd; clone number: OTI6H8) or anti-Mouse Ki-67 monoclonal antibody (1:2500 dilution; Beijing Zhongshan Golden Bridge Biotechnology; clone number: UMAB107).

Techniques:

Journal: Diabetes

Article Title: Transient PAX8 Expression in Islets During Pregnancy Correlates With β-Cell Survival, Revealing a Novel Candidate Gene in Gestational Diabetes Mellitus.

doi: 10.2337/db18-0285

Figure Lengend Snippet: Figure 1. Pax8 is transiently expressed in pancreatic islets during gestation. (A-C) Determination of Pax8 (A), Tph1 (B) and Pdx1 (C) mRNA levels measured by real time RT-PCR in murine islets at different stages of pregnancy. n=4-5 animals per time point were used. (D-F) Time course determination of PAX8 (D), TPH1 (E) and PDX1 (F) mRNA levels measured by real time RT-PCR in human islets treated daily with prolactin (200 ng/ml). n=8 per group at times 0 h, 24 h, 48, 72 h and n=7 in time point 96 h in panel D. n=4 per time point in panel E. n=5 per in times 0 h, 24 h, 48, 72 h and n=3 in time point 96 h in panel F. (G) Determination of proliferation by BrDU incorporation in human islets at different prolactin treatment time points. BrDU was added to the culture 24 hours prior processing the samples n=5-6 islet preparations were analyzed per time point. NP: Non-pregnant. Data are represented as the mean ± SD of non-pregnant females or 0 hours of treatment. * p< 0.05 compared to non-pregnant mice or time 0 on human islets.

Article Snippet: Generation of PAX8 lentiviruses and transduction The human and murine PAX8 wild type cDNAs (Origene, MD, USA) were subcloned into the pHRSIN DUAL-GFP easy vector, kindly provided by Dr. Pintor-Toro (CABIMER, Seville, Spain).

Techniques: Quantitative RT-PCR, BrdU Incorporation Assay

Journal: Diabetes

Article Title: Transient PAX8 Expression in Islets During Pregnancy Correlates With β-Cell Survival, Revealing a Novel Candidate Gene in Gestational Diabetes Mellitus.

doi: 10.2337/db18-0285

Figure Lengend Snippet: Figure 2. PAX8 targets different genetic pathways in human and mouse islet that converged in improved islet survival. Human and murine pancreatic islets were transduced with mock (GFP control), mouse Pax8 (Pax8) or human PAX8 (PAX8) lentiviral vectors. (A) Flow cytometry analysis according to size (FSC) and granularity (SSC). FSC: forward scattered light. SSC: side scattered light. (B) Flow cytometry histograms showing the number of events plotted against GFP fluorescence. The line indicates the threshold for positivity. (C-D) DNA microarray profiling was performed and the Principal component analysis (PCA) was applied on the data derived from (C) murine and (D) human islets. Each point corresponds to the PCA analysis of one biological replicate. n=3 independent replicates. (E) Heat map depicting the relative expression levels of shared significantly modulated genes as compared to control in murine and human pancreatic islets. (F-G) Comparison of genetic pathways significantly altered by the overexpression of the murine Pax8 (F) or the human PAX8 (G) as compared to mock-transduced islets using the TAC platform. (H) Determination of gene expression on murine pancreatic islets infected with Mock, Pax8 or PAX8. n=3 biological replicates per experimental group. (I) Determination of metabolic activity using the MTT assay in transduced murine islets. n=4 per condition. (J) Insulin secretion of transduced murine islets was assessed

Article Snippet: Generation of PAX8 lentiviruses and transduction The human and murine PAX8 wild type cDNAs (Origene, MD, USA) were subcloned into the pHRSIN DUAL-GFP easy vector, kindly provided by Dr. Pintor-Toro (CABIMER, Seville, Spain).

Techniques: Transduction, Control, Flow Cytometry, Fluorescence, Microarray, Derivative Assay, Expressing, Comparison, Over Expression, Gene Expression, Infection, Activity Assay, MTT Assay

Journal: Diabetes

Article Title: Transient PAX8 Expression in Islets During Pregnancy Correlates With β-Cell Survival, Revealing a Novel Candidate Gene in Gestational Diabetes Mellitus.

doi: 10.2337/db18-0285

Figure Lengend Snippet: Figure 3. The PAX8-T356M and P25R mutations on PAX8 are found in two patients that developed GDM and both mutations exhibit compromised transcriptional activity. (A) Family pedigree 1 (T356M) with gestational as well as T2DM diabetes mellitus and hypothyroidism in consecutive generations. Circles indicate females and squares indicate males. (B) Sequencing chromatogram of T356M mutation in the index patient of pedigree 1 and a wild type sequence in a healthy relative. (C) Schematic representation of PAX8 protein with annotated known mutations including T356M and P25R. (D-F) Oral glucose tolerance test during pregnancy (week 24-28) on the index patient harboring T356M. Levels above the threshold of NDGG and/or CC diagnosis for GDM are highlighted in red. (D) T356M, second pregnancy. (E) T356M, third pregnancy. (F) T356M, fourth pregnancy. (G) Family pedigree 2 (P25R) with GDM and hypothyroidism. (H) Oral glucose tolerance test during the first pregnancy on index patient of pedigree 2 harboring P25R. (I-J) MCF7 cells were transfected with an empty pCDNA3.1, wild type (PAX8Wt) pCDNA3.1, PAX8 T356M pCDNA3.1 (T356M) or PAX8 P25R pCDNA3.1 (P25R) plasmid. Transfected cells were then incubated with 80 μg/mL cycloheximide (CHX) for the indicated time. Transfected PAX8 and endogenous β-actin protein levels were determined by standard immunoblotting. n=3 independent experiments performed in triplicate. (I)

Article Snippet: Generation of PAX8 lentiviruses and transduction The human and murine PAX8 wild type cDNAs (Origene, MD, USA) were subcloned into the pHRSIN DUAL-GFP easy vector, kindly provided by Dr. Pintor-Toro (CABIMER, Seville, Spain).

Techniques: Activity Assay, Sequencing, Mutagenesis, Biomarker Discovery, Transfection, Plasmid Preparation, Incubation, Western Blot