stat5b proteins  (Thermo Fisher)

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: STAT5B mutations at Y665 in the mouse genome. A Schematic illustration of the STAT5B protein domains. Amino acid locations of the variants are highlighted. B Three-dimensional model of STAT5B protein structure presenting the location of the Y665 residue mutated in the mouse genome. C Sanger sequencing chromatograms showing Y665 (wild-type, WT) and the introduction of SNPs resulting in the two missense mutations, Stat5b Y665 F (Y665F) and Stat5b Y665H (Y665H) mutants. The red shade indicates the altered codons converting Y665 to Y665F and Y665H, respectively. D Number and percentage of offspring from intercrossed heterozygote parents (Y665H, n = 301; Y665F, n = 543). The statistical relationship between the percentage and the expected Mendelian ratios was assessed using a Chi-square test. E Dot plot presenting the weight of individual mice in each group from 6, 7, 8 and 9 weeks of age. Wild-type mice are from litters weaned from heterozygous Stat5b Y665H (gray) and Stat5b Y665F (black) dams. Results are presented as the means ± SEM of independent biological replicates (Y665, n = 13; Y665H, n = 6; Y665F, n = 17). A two-way ANOVA followed by Tukey’s multiple comparisons test was used to evaluate the statistical significance of differences between groups. ** p < 0.001, **** p < 0.0001

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: Residue, Sequencing

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: Impaired alveolar development in STAT5B Y665 mutant mice at day 18.5 of pregnancy. A Sections of mammary tissue stained with hematoxylin and eosin at day 18.5 of pregnancy with high magnification (x400, Scale bars, 100 μm). The arrow and Asterix indicates alveoli and lumen. Y665, n = 3; Y665H, n = 3; Y665F, n = 4. 50–100 of the alveolar architecture on each slide was quantitatively analyzed using the ImageJ software, with the results visualized through dot plots. (a) milk-secreting alveoli; (b) alveolar lumen; (c) alveolar epithelial cells. B , D and E Heatmaps showing fold activity of significantly regulated genes of milk proteins in red, enzymes in green, fat metabolism in gray, membrane transporters in blue, transcription factors in black (B), cell marker genes (D) and skin-related genes (E) in STAT5B Y665H and STAT5B Y665F mutants compared to WT (RNA-seq; Y665, n = 3; Y665H, n = 3; Y665F, n = 4). C Dot plots with the normalized read counts from RNA-seq to mRNA levels of Stat5 genes controlled by autoregulatory enhancer. Results are shown as the means ± SEM of independent biological replicates (Y665, n = 3; Y665H, n = 3). The Benjamini-Hochberg adjusted p -value was used for significance. *** p < 0.0001, **** p < 0.0001

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: Mutagenesis, Staining, Software, Activity Assay, Membrane, Marker, RNA Sequencing

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: Nuclear localization of STAT5A and STAT5B in STAT5B Y665 mutant mice. Staining of STAT5A and STAT5B proteins at day 18.5 of the first pregnancy with high magnification (x10 and x400, Scale bars, 100 μm). The arrow indicates alveolar lumen ( a ), alveolar epithelial cells ( b ), fibroblasts ( c ) and ( d ) fat

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: Mutagenesis, Staining

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: Genomic features of the genes regulated by STAT5B Y665 mutations. A Coverage plots displaying the patterns of H3K27ac marks and STAT5B and STAT5A binding on 440 mammary-specific super-enhancers at day 18.5 of pregnancy. Black, Y665; blue, Y665H; red, Y665F. B Binding of STAT5B and NFIB, H3K27ac, and Pol II at gene loci of milk protein genes controlled by mammary-specific super-enhancers, transporter genes, and skin genes in mammary tissue of wild type (Y665) and Stat5b Y665H (Y665H) mice at day 18.5 of pregnancy. C STAT5B and NFIB binding, H3K27ac and Pol II loading at gene loci of milk protein genes controlled by mammary-specific super-enhancers, transporter gene and skin gene in mammary tissue of Y665 and Stat5b Y665F (Y665F) mice. Y665, n = 2; Y665H, n = 2; Y665F, n = 2

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: Binding Assay

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: Alveolar development of STAT5B Y665F mutants at mid-pregnancy. A Sections of mammary tissue stained with hematoxylin and eosin at day 13.5 of pregnancy with high magnification (x400, Scale bars, 100 μm). The arrow indicates alveoli. Y665, n = 3; Y665F, n = 4. The alveolar architecture was quantitatively analyzed using the ImageJ software, with the results visualized through dot plots. (a) milk-secreting alveoli; (b) alveolar lumen; (c) alveolar epithelial cells. B Bar graphs showing fold activity of genes of milk proteins, transporters and transcription factors in STAT5B Y665F (Y665F) mice compared to wild type (Y665) (RAN-seq; Y665, n = 3; Y665F, n = 4). C Heatmaps presenting fold activity of milk protein genes in red, transporter genes in blue, transcription factors in black (left) and skin genes (right) in STAT5B Y665F mice at day 13.5 of pregnancy (presented as p13) compared to WT at day 18.5 of pregnancy (presented as p18) (RAN-seq; Y665 at p18, n = 3; Y665F at p13, n = 4). D Binding of STAT5B and NFIB, H3K27ac, and Pol II at gene loci of milk protein genes controlled by mammary-specific super-enhancers in mammary tissue of Y665 and Y665F mice at day 13.5 of pregnancy. Y665, n = 2; Y665F, n = 2

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: Staining, Software, Activity Assay, Binding Assay

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: Partial activation of mammary function and lactation in STAT5B Y665H mice after the second pregnancy. A Body weight of pups at day ten of lactation (L10) after the second pregnancy of homozygous Stat5b Y665H mice and after the first pregnancy of heterozygous Stat5b Y665H mice. The weight of pups from 3–4 females was measured and normalized by the number of pups. A t -test was utilized to evaluate the statistical significance between pups from homozygous females at second pregnancy and heterozygous females at first pregnancy. Homozygous Stat5b Y665H mice, n = 4 cages; heterozygous Stat5b Y665H mice, n = 3 cages. * p < 0.05. B Expression of representative mammary genes was measured in mammary tissue of Y665 (WT) mice collected at day 10 of lactation (L10), and of STAT5B Y665H mice at L10 after the second pregnancy by RNA-seq (Y665, n = 3; Y665H, n = 4). The p18 data (Fig. ) is presented alongside the L10 data to highlight the relative reduction in gene expression between p18 and L10 in Y665H mice. C Binding of STAT5A and STAT5B, H3K27ac, and Pol II at Stat5 and Wap super-enhancer loci controlled by prolactin in mammary tissue of Y665 and Y665H mice at day 10 of lactation. The Socs2 locus was presented as a ChIP-seq control. Y665, n = 2; Y665H, n = 2. D Sections of mammary tissue stained with hematoxylin and eosin at day 10 of lactation after the second pregnancy with high magnification (x400, Scale bars, 100 μm). The arrow and Asterix indicates alveoli and lumen. Y665, n = 1; Y665H, n = 3. The alveolar architecture was quantitatively analyzed using the ImageJ software, with the results visualized through dot plots. (a) milk-secreting alveoli; (b) alveolar lumen; (c) alveolar epithelial cells

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: Activation Assay, Expressing, RNA Sequencing, Gene Expression, Binding Assay, ChIP-sequencing, Control, Staining, Software

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: Regulation of key fat metabolic genes. A-B Dot plots with the normalized read counts from RNA-seq to mRNA levels of Olah and Fabp3 genes at virgin and L10 of wild type mice (A) and at p18 of wild type and Y665 mutant mice (B). Results are shown as the means ± SEM of independent biological replicates (WT at virgin, n = 3; WT at L10, n = 4; Y665 at p18, n = 3; Y665H at p18, n = 3). p -values are from 2-way ANOVA followed by Sidak’s multiple comparisons test between WT and mutants. ** p < 0.001, **** p < 0.0001. C Binding of SAT5A and STAT5B, H3K27ac, and Pol II at Olah and Fabp3 gene loci controlled by prolactin in mammary tissue of Y665 and Y665H mice at day 10 of lactation. Y665, n = 2; Y665H, n = 2

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: RNA Sequencing, Mutagenesis, Binding Assay

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Disease-Associated Mutations of the STAT5B SH2 Domain Regulate Cytokine-Driven Enhancer Function and Mammary Development

doi: 10.1007/s10911-025-09582-8

Figure Lengend Snippet: Dimerization of STAT5A and STAT5B protein. A Strategy of co-ChIP-seq. STAT5A bound chromatin from mammary tissue of WT mice at day 1 of lactation (L1) was precipitated using STAT5A antibody and eluted from Ig beads. Secondly, STAT5B bound chromatin was precipitated using STAT5B and eluted from the beads. STAT5A motifs from 1st ChIP and STAT5B motifs from 2nd ChIP were identified by sequencing. B Co-ChIP profile at the representative milk protein Wap gene and common STAT target Socs2 gene. C STAT5A and STAT5B binding at gene loci of milk protein Wap gene and common STAT target Socs2 gene in the mammary gland of Y665 (WT) and Y665H mice at day 18.5 of pregnancy (p18). D STAT5A and STAT5B binding in the mammary gland of Y665 (WT) and Y665F mice at day 13.5 of pregnancy (p13)

Article Snippet: The expression of STAT5A and STAT5B proteins was detected by overnight incubation with primary antibodies: STAT5A (1:200, Santa Cruz Biotechnology, sc-271542) and STAT5B (1:200, R&D Systems, AF1584) at room temperature.

Techniques: ChIP-sequencing, Sequencing, Binding Assay

Journal: Nutrients

Article Title: Lactation Activity and Mechanism of Milk-Protein Synthesis by Peptides from Oyster Hydrolysates

doi: 10.3390/nu14091786

Figure Lengend Snippet: Primer sequences for qRT-PCR analysis.

Article Snippet: AKT1, α s1 -casein (CSN1S1) and Cyclin D1 were sourced from Proteintech (Rosemont, IL, USA); P-AKT and p-S6K1 were obtained from Cell Signaling (Danfoss, MA, USA); S6K1, mTOR, P-mTOR, κ-casein (CSN3), β-lactoglobulin (BLG), β-casein (CSN2), prolactin (PRL), prolactin receptor (PRLR), STAT5a, STAT5b and β-actin were obtained from Affinity biosciences, OH, USA; and casein enzymatic hydrolysate was obtained from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China).

Techniques:

Journal: Radiation research

Article Title: Transcript Analysis for Internal Biodosimetry Using Peripheral Blood from Neuroblastoma Patients Treated with 131 I-mIBG, a Targeted Radionuclide

doi: 10.1667/RR14263.1

Figure Lengend Snippet: Selected Transcripts of Interest

Article Snippet: The DNA repair domain was represented by XPC and DDB2 . table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Gene Name Primer no. GAPDH Glyceraldehyde 3-phosphate dehydrogenase H202758991_g1 CDKN1A Cyclin-dependent kinase inhibitor 1A (p21) Hs0035578_m1 FDXR Ferrodixin reductase Hs00244586_m1 GADD45A Growth arrest and DNA-damage-inducible alpha Hs00169255_m1 DDB2 Damage-specific DNA binding protein 2 (XPE) Hs03044953_m1 XPC Xeroderma pigmentosum, complementation group C Hs00190295_m1 MDM2 E3 ubiquitin protein ligase Hs00234753_m1 BCL2 B-cell CLL/lymphoma 2 Hs99999018_m1 BCLXL BCL2-like 1 Hs00236329_m1 BAX BCL2-associated X protein Hs99990001_m1 STAT5B Signal transducer and activator of transcription 5B Hs00273500_m1 Open in a separate window Selected Transcripts of Interest Generation of cDNA and Real-Time PCR For RT-PCR analysis, 200 ng of RNA was converted to single-strand cDNA using the High Capacity cDNA archive kit (Applied Biosystems ® , Foster City, CA).

Techniques: Binding Assay

Journal: Radiation research

Article Title: Transcript Analysis for Internal Biodosimetry Using Peripheral Blood from Neuroblastoma Patients Treated with 131 I-mIBG, a Targeted Radionuclide

doi: 10.1667/RR14263.1

Figure Lengend Snippet: Box-whisker plots show the modulation of selected transcripts over time. This collection of plots shows the changes in gene expression at each time interval after 131I-mIBG treatment. The x-axis shows the time points sampled (0–96 h), while the y-axis shows the transformed log2 fold change of expression levels. The transcripts BAX, CDKN1A, DDB2, FDXR, GADD45A, MDM2, STAT5B and XPC show time-dependent upregulation while BCLXL shows time-dependent downregulation (paired Student's t test vs. 0 h after treatment, ***P < 0.001). BCL2 did not have a time-dependent response to radiation.

Article Snippet: The DNA repair domain was represented by XPC and DDB2 . table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Gene Name Primer no. GAPDH Glyceraldehyde 3-phosphate dehydrogenase H202758991_g1 CDKN1A Cyclin-dependent kinase inhibitor 1A (p21) Hs0035578_m1 FDXR Ferrodixin reductase Hs00244586_m1 GADD45A Growth arrest and DNA-damage-inducible alpha Hs00169255_m1 DDB2 Damage-specific DNA binding protein 2 (XPE) Hs03044953_m1 XPC Xeroderma pigmentosum, complementation group C Hs00190295_m1 MDM2 E3 ubiquitin protein ligase Hs00234753_m1 BCL2 B-cell CLL/lymphoma 2 Hs99999018_m1 BCLXL BCL2-like 1 Hs00236329_m1 BAX BCL2-associated X protein Hs99990001_m1 STAT5B Signal transducer and activator of transcription 5B Hs00273500_m1 Open in a separate window Selected Transcripts of Interest Generation of cDNA and Real-Time PCR For RT-PCR analysis, 200 ng of RNA was converted to single-strand cDNA using the High Capacity cDNA archive kit (Applied Biosystems ® , Foster City, CA).

Techniques: Whisker Assay, Expressing, Transformation Assay

Journal: Radiation research

Article Title: Transcript Analysis for Internal Biodosimetry Using Peripheral Blood from Neuroblastoma Patients Treated with 131 I-mIBG, a Targeted Radionuclide

doi: 10.1667/RR14263.1

Figure Lengend Snippet: Multiple Regression Analysis Results for Gene Expression Based on Dose and Time

Article Snippet: The DNA repair domain was represented by XPC and DDB2 . table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Gene Name Primer no. GAPDH Glyceraldehyde 3-phosphate dehydrogenase H202758991_g1 CDKN1A Cyclin-dependent kinase inhibitor 1A (p21) Hs0035578_m1 FDXR Ferrodixin reductase Hs00244586_m1 GADD45A Growth arrest and DNA-damage-inducible alpha Hs00169255_m1 DDB2 Damage-specific DNA binding protein 2 (XPE) Hs03044953_m1 XPC Xeroderma pigmentosum, complementation group C Hs00190295_m1 MDM2 E3 ubiquitin protein ligase Hs00234753_m1 BCL2 B-cell CLL/lymphoma 2 Hs99999018_m1 BCLXL BCL2-like 1 Hs00236329_m1 BAX BCL2-associated X protein Hs99990001_m1 STAT5B Signal transducer and activator of transcription 5B Hs00273500_m1 Open in a separate window Selected Transcripts of Interest Generation of cDNA and Real-Time PCR For RT-PCR analysis, 200 ng of RNA was converted to single-strand cDNA using the High Capacity cDNA archive kit (Applied Biosystems ® , Foster City, CA).

Techniques: Expressing