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: Nature Communications

Article Title: SOX17-positive rete testis epithelium is required for Sertoli valve formation and normal spermiogenesis in the male mouse

doi: 10.1038/s41467-022-35465-1

Figure Lengend Snippet: a , b Schematic illustration of the experiment and Uniform Manifold Approximation and Projection (UMAP) plot of cells isolated from the proximal part of the testis of P14 Sox17- cKO and controls, showing twenty clusters ( a ) and the distribution of cells from each sample ( b ). Control n = 2, Sox17- cKO n = 2. c UMAP plot showing the expression level of Sox9, Pax8, and Sox17 in the control samples. The right two panels show the expression of each gene in the extracted clusters of the RT (cluster 16) and Sertoli cells (clusters 1, 2, 10, 18). d Dot plots representing the expression levels of marker genes for Sertoli cells ( Amh ) and RT epithelial cells ( Krt8, Pax8 ) together with their common marker genes ( Nr5a1, Gata4, Wt1, Sox9 ) in the clusters expressing Sox9 shown in ( c ) (clusters 1, 2, 10, 16, 18). e A violin plot showing the expression level of Sox17 in each cluster, together with Pecam1 (vascular endothelial cell marker) and Pax8 (RT marker). “ n ” represents the number of biological replicates.

Article Snippet: The sections were incubated overnight at 4 °C with the following primary antibodies [format: host anti-protein (company, catalog number, dilution)]: mouse monoclonal anti-ace-TUB (Sigma, T6793, 1:200), mouse monoclonal anti-αSMA/ACTA2 (Sigma, A5228, 1:500), goat polyclonal anti-AMH (Santa Cruz, sc6886, 1:200), mouse monoclonal anti-CDH1 (BD Transduction lab, 610181, 1:400), goad polyclonal anti-c-KIT (R&D systems, AF1356, 1:200), rabbit monoclonal anti-EPCAM (Abcam, ab32392, 1:100), goat polyclonal anti-GATA4 (Santa Cruz, sc1237, 1:200), mouse monoclonal anti-GATA4 (Santa Cruz, sc25310, 1:100), rabbit polyclonal anti-GFP (MBL, 598, 1:200), goat polyclonal GFRα1 (R&D systems, AF560, 1:100), rabbit polyclonal anti-HSP70 (Abcam, ab79852, 1:1000), rabbit polyclonal anti-KI67 (Abcam, ab15580, 1:400), rabbit monoclonal anti-KRT8 (Abcam, ab53280, 1:200), rabbit polyclonal LAMININ (Abcam, ab11575, 1:200), rabbit monoclonal anti-p-AKT (Cell Signaling, 4060, 1:100), mouse monoclonal anti-PAX8 (Abcam, ab53490, 1:100), rat monoclonal PECAM1 (Invitrogen, 14-0311-82, 1:100), rabbit polyclonal anti-PLZF (Santa Cruz, sc22839, 1:200), mouse monoclonal SCP3 (Santa Cruz, sc74569, 1:500), goat polyclonal anti-SOX17 (R&D systems, AF1924, 1:200), rabbit polyclonal anti-SOX9 (Millipore, AB5535, 1:400), rabbit polyclonal anti-STAR (Cell Signaling, 8449, 1:100), rabbit polyclonal anti-VASA/MVH (Abcam, ab13840, 1:10000).

Techniques: Isolation, Expressing, Marker

Journal: BMC Molecular Biology

Article Title: Pax8 modulates the expression of Wnt4 that is necessary for the maintenance of the epithelial phenotype of thyroid cells

doi: 10.1186/1471-2199-15-21

Figure Lengend Snippet: Wnt4 is localized in the cytoplasmic compartment of FRTL-5 cells and its expression is modulated by TSH. A) Wnt4 expression is modulated by TSH in FRTL-5 thyroid cells. FRTL-5 cells were cultured in regular medium or in starvation medium for 4 days (0.2% CS) and treated with 1 mU/ml of TSH for 24 h, and analyzed by immunofluorescence to detect Pax8 and Wnt4 proteins with specific antibodies (see Methods). B) TSH stimulation of Wnt4 expression is mediated by Pax8. FRTL-5 cells were cultured in regular medium and transfected with 100 nM of siPax8 or siCtrl-. Seven hours later, cells were starved in 0.2% CS medium for 24 h and treated with 1 mU/ml of TSH for 3 h and analyzed by qRT-PCR to measure the expression levels of Pax8 (black bars) and Wnt4 (gray bars) mRNAs. Data are from three independent experiments, each performed in duplicate and are expressed as the mean ± SD (P < 0,1).

Article Snippet: Briefly, the cells were subsequently blocked with washing buffer containing 0.5% BSA for 1 h and treated with rabbit policlonal anti-WNT4 (Wnt4 antibody, Novus) O/N and then with Alexa Fluor ® 594-conjugated secondary antibody (Invitrogen) for 30 min, followed by treatment with mouse monoclonal anti-PAX8 (BIOCARE MEDICAL) for 1 h and finally by an Alexa Fluor ® 488-conjugated secondary antibody (Invitrogen).

Techniques: Expressing, Cell Culture, Immunofluorescence, Transfection, Quantitative RT-PCR

Journal: BMC Molecular Biology

Article Title: Pax8 modulates the expression of Wnt4 that is necessary for the maintenance of the epithelial phenotype of thyroid cells

doi: 10.1186/1471-2199-15-21

Figure Lengend Snippet: Pax8 activates transcription from the Wnt4 5′-flanking region. A) Transcriptional activity of Wnt4 5′-flanking genomic region. The deletion reporter constructs were transfected into FRTL-5 and HeLa cells and the luciferase activity was determined. All the constructs have the highest activity in thyroid cells and only the 190Wnt4LUC construct shows a strongly reduced activity. Data are expressed as the mean ± SD (P < 0,0001); B) Pax8 activates transcription from the Wnt4 promoter region. HeLa cells were transiently transfected with 300Wnt4LUC and 190Wnt4LUC reporter constructs alone and in combination with increasing concentration (100 and 200 ng) of the expression vector encoding Pax8 (CMV5-Pax8). The transcriptional activity was determined 48 h after transfection as the firefly over renilla luciferase activity. Data are expressed as fold induction over the transcription obtained with 300Wnt4LUC and 190Wnt4LUC, whose values were set at 1. Data are from three independent experiments, each performed in duplicate and are expressed as the mean ± SD (P < 0,005); C) Pax8 binds the Wnt4 promoter region in vivo. Chromatin extracted from cross-linked FRTL-5 cells was immunoprecipitated with an unrelated antibody or for Pax8. The immunoprecipitates were analyzed by PCR with oligonucleotides able to amplify the region between -300 bp and -190 bp of the 5′flanking region of the Wnt4 gene.

Article Snippet: Briefly, the cells were subsequently blocked with washing buffer containing 0.5% BSA for 1 h and treated with rabbit policlonal anti-WNT4 (Wnt4 antibody, Novus) O/N and then with Alexa Fluor ® 594-conjugated secondary antibody (Invitrogen) for 30 min, followed by treatment with mouse monoclonal anti-PAX8 (BIOCARE MEDICAL) for 1 h and finally by an Alexa Fluor ® 488-conjugated secondary antibody (Invitrogen).

Techniques: Activity Assay, Construct, Transfection, Luciferase, Concentration Assay, Expressing, Plasmid Preparation, In Vivo, Immunoprecipitation

Journal: BMC Molecular Biology

Article Title: Pax8 modulates the expression of Wnt4 that is necessary for the maintenance of the epithelial phenotype of thyroid cells

doi: 10.1186/1471-2199-15-21

Figure Lengend Snippet: Wnt4 5′-flanking region contains a binding site for Pax8. A) Graphic output of the sequence analysis showing the conservation in different species of the core consensus sequences (in bold) of Pax8 binding sites. The sequence alignment was obtained using whole genome comparative analysis of the VISTA browser; B) Electromobility shift assays for Pax8 binding to putative recognition motifs in the Wnt4 promoter. Left panel, 32 P-labeled oligo probes A and B were challenged with total protein extracts prepared from FRTL-5 cells (lane 2). Protein extracts of HeLa (lane 3) and Pax8-transfected HeLa cells (lane 4) were used as negative and positive control, respectively. Middle panel, the specificity of the complex observed with the FRTL-5 extract and probe A was tested by competition analysis, using increasing amount (from 25 to 100-fold molar excess) of unlabeled wild-type oligo A or mutated in the core sequence. Right panel, the FRTL-5 protein extract was incubated with the antibody against Pax8 or tubulin (as negative control), in a supershift EMSA with labeled probe A; C) Pax8 binding site A is necessary for the transcriptional activity of Wnt4 promoter. The deletion constructs 300Wnt4LUC and 260Wnt4LUC were transfected into FRTL-5 thyroid cells and the luciferase activity was determined. Data are from three independent experiments, each performed in duplicate and are expressed as the mean ± SD (P < 0,05).

Article Snippet: Briefly, the cells were subsequently blocked with washing buffer containing 0.5% BSA for 1 h and treated with rabbit policlonal anti-WNT4 (Wnt4 antibody, Novus) O/N and then with Alexa Fluor ® 594-conjugated secondary antibody (Invitrogen) for 30 min, followed by treatment with mouse monoclonal anti-PAX8 (BIOCARE MEDICAL) for 1 h and finally by an Alexa Fluor ® 488-conjugated secondary antibody (Invitrogen).

Techniques: Binding Assay, Sequencing, Labeling, Transfection, Positive Control, Incubation, Negative Control, Activity Assay, Construct, Luciferase

Journal: BMC Molecular Biology

Article Title: Pax8 modulates the expression of Wnt4 that is necessary for the maintenance of the epithelial phenotype of thyroid cells

doi: 10.1186/1471-2199-15-21

Figure Lengend Snippet: Pax8 and Wnt4 expression in human thyroid cancer cell lines. Wnt4 and Pax8 expression was measured by qRT-PCR in four human thyroid cancer cell lines: WRO from follicular thyroid cancer, Cal62 from anaplastic thyroid carcinoma, FB2 and BCPAP from papillary thyroid carcinoma. RNA from six non-pathological thyroids (N.T.) was used as control. Quantitative real-time PCR was carried out in triplicate as described in Materials and Methods. Glyceraldehyde 3-phosphate dehydrogenase was used as reference gene; results are reported as 2 -∆Ct . Data are expressed as the mean ± SD (P < 0,05). The bottom panel shows the total protein extracts of FRTL-5 cells (used as normal thyroid) and human thyroid cancer cells WRO, Cal62, FB2 and BCPAP separated on SDS-PAGE and subjected to Western blot analysis with specific antibodies as indicated in the panel. The hybridization with GAPDH assessed the protein uniform loading integrity. Fold represents the quantification of protein levels normalized with GAPDH.

Article Snippet: Briefly, the cells were subsequently blocked with washing buffer containing 0.5% BSA for 1 h and treated with rabbit policlonal anti-WNT4 (Wnt4 antibody, Novus) O/N and then with Alexa Fluor ® 594-conjugated secondary antibody (Invitrogen) for 30 min, followed by treatment with mouse monoclonal anti-PAX8 (BIOCARE MEDICAL) for 1 h and finally by an Alexa Fluor ® 488-conjugated secondary antibody (Invitrogen).

Techniques: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, SDS Page, Western Blot, Hybridization