Journal: Human Molecular Genetics

Article Title: Inherited thrombocytopenia associated with mutation of UDP-galactose-4-epimerase ( GALE )

doi: 10.1093/hmg/ddy334

Figure Lengend Snippet: Inherited thrombocytopenia in family AH; GALE conservation, structure and mutations. (A) Six individuals in four-generation family AH presented with severe thrombocytopenia (black symbols). Current ages in years of individuals are listed below symbols. V indicates variant allele GALE p.R51W (c.C151T); N indicates reference allele. Asterisks indicate individuals evaluated by whole-exome sequencing. (B) Bone marrow aspirate treated with Wright-Giemsa stain from affected family member IV.1. Arrow indicates a megakaryocyte with hypogranular cytoplasm and hypolobated nuclei. (C) Protein sequence alignment from multiple species of the GALE region including Arg51 (red text) showing evolutionary conservation. Gold indicates complete conservation through S. cerevisiae. (D) GALE crystal structure (PDB ID:1HZJ), viewed in PyMOL (72). Left: GALE homodimer (green and gold), highlighting Arg51 (magenta), Asn340 (orange), UDP-N-acetylgalactosamine (blue), NAD+ cofactor (red), C-terminal end of GALE (yellow). Right: close-up view of Arg51 near the C-terminal end of GALE. Dotted yellow line indicates the 3-Å distance between Arg51 and Asn340. (E) Galactosemia-associated mutations of GALE reported in the literature. GALE domains are the substrate-binding site (gold), NAD+-binding site (orange) and catalytic sites (red dots at residues 132, 157, 307). Asterisks indicate mutations reported as homozygotes; others were reported as compound heterozygotes. GALE p.V94M (red text) is associated with severe generalized galactosemia. GALE p.R51W is shown in bold.

Article Snippet: TaqMan probes for GALE (Hs00166181_m1) and control GAPDH were used.

Techniques: Variant Assay, Sequencing, Giemsa Stain, Binding Assay

Journal: Human Molecular Genetics

Article Title: Inherited thrombocytopenia associated with mutation of UDP-galactose-4-epimerase ( GALE )

doi: 10.1093/hmg/ddy334

Figure Lengend Snippet: Enzyme activity and differential scanning calorimetry. Enzyme assays in figures (A) and (B) were performed with 10-ng protein. Substrates were (A) UDP-galactose or (B) UDP-N-acetylgalactosamine. Enzyme activities were measured as fraction product (Supplementary Material, Table S2) and were normalized to wildtype protein measurements. The mean values of normalized enzyme activities were plotted + standard deviation for each allele. Experiments were performed in at least triplicate (23,28,29). (C) Representative endotherm plots for each GALE allele. Endotherm data was normalized for protein concentration, with intrascan baseline correction. Black indicates wildtype protein (normal), red indicates GALE p.R51W, purple indicates GALE p.V94M and orange indicates GALE p.D103G. (D) Size distributions of megakaryocyte colonies after treatment of CD34+ hematopoietic progenitor cells with control or anti-GALE shRNA probes. Cells were plated in MegaCult collagen-based media. Slides were fixed, stained and classified by colony size. Data represents two independent experiments, each with plating and counting in duplicate.

Article Snippet: TaqMan probes for GALE (Hs00166181_m1) and control GAPDH were used.

Techniques: Activity Assay, Standard Deviation, Protein Concentration, shRNA, Staining

Journal: Human Molecular Genetics

Article Title: Inherited thrombocytopenia associated with mutation of UDP-galactose-4-epimerase ( GALE )

doi: 10.1093/hmg/ddy334

Figure Lengend Snippet: N-linked glycosylation pathway and thrombocytopenia. Mutations in critical steps of the N-linked glycan biosynthesis pathway are associated with thrombocytopenia. Glycan biosynthesis begins in the cytoplasm, with subsequent modifications in the endoplastic reticulum and the Golgi. Membrane transporters including SLC35A1, SLC35A2 and SLC35A3 allow building-block sugars to enter the Golgi. Enzymes in the pathway, including B4GALT1, control the levels of these sugars, and add or subtract sugars from the glycan chain. GALE is responsible for interconversion of these sugars in the cytoplasm, in preparation for their entry into the Golgi and integration into glycan chains. Mutations in this pathway cause CDGs. Two forms of CDG include thrombocytopenia among presenting symptoms: CDG-IId, caused by mutation of B4GALT1, and CDG-IIf, caused by mutation of SLC35A1. Genes responsible for steps in the pathway are indicated in green boxes, with the CDG disorder under the gene name. Steps in the pathway with mutations associated with thrombocytopenia are indicated by red Xs. Figure is adapted from Freeze 2006 (73).

Article Snippet: TaqMan probes for GALE (Hs00166181_m1) and control GAPDH were used.

Techniques: Blocking Assay, Mutagenesis

Journal: The Journal of Biological Chemistry

Article Title: Redox imbalance induces remodeling of glucose metabolism in Rhipicephalus microplus embryonic cell line

doi: 10.1016/j.jbc.2022.101599

Figure Lengend Snippet: Oxidative challenge upregulates transcriptional levels of glycogen synthesis components in BME26 cells. Transcription of ( A ) glycogen synthase (GS), ( B ) glycogen synthase kinase 3 β (GSK3β), ( C ) phosphoglucomutase (PGM), and ( D ) glycogen debranching enzyme (GDE) in Rhipicephalus microplus embryonic cell line (BME26) was measured 2 and 24 h after addition of H 2 O 2 (2 or 4 mM). E , diagram of glycogen metabolism depicting pathways involved in glycogen synthesis and degradation. Experiments were performed in three independent biological samples with three experimental replicates each, where ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, compared with the control; and # p < 0.05, #### p < 0.0001, compared between the groups, in Tukey’s multiple comparisons test.

Article Snippet: Cells were kept for 1 h in blocking solution (1% bovine serum albumin in PBS), followed by a 16-h incubation at 4 to 8 °C with the respective primary antibody, anti-Glycogen Synthase (GS, total) (Cell signaling, #3893) or anti-Glycogen Synthase phosphorylated at Ser641 (pGS, inhibited) (Cell signaling, #3891), at 1:200 dilution in blocking solution.

Techniques: Control

Journal: The Journal of Biological Chemistry

Article Title: Redox imbalance induces remodeling of glucose metabolism in Rhipicephalus microplus embryonic cell line

doi: 10.1016/j.jbc.2022.101599

Figure Lengend Snippet: Glycogen metabolism regulation in BME26 cells upon H 2 O 2 challenge. H 2 O 2 -tolerant BME26 cells increased glycogen synthase protein expression 2 h after challenge with 2 or 4 mM H 2 O 2 . A , immunolocalization ( red signal ) of phosphorylated Ser 641 glycogen synthase (pGS) or total GS, as well as nuclei staining ( blue signal ), was performed after 2 or 4 mM H 2 O 2 challenge, for 2 and 24 h. Images were captured in a confocal laser scanning microscope, LSM 710, Zeiss. The scale bar represents 10 μm. B – D , quantitative analysis of GS ( B ), pGS ( C ), and active GS ( D ). E , periodic acid–Schiff staining of BME26 cells after treatment with 2 and 4 mM H 2 O 2 for 2 and 24 h. Blue arrows indicate diffuse staining and red arrows indicate perinuclear staining. Light microscope images were captured in bright field using Axio Scope.A1, Zeiss polarized light microscope through the Blue Zeiss software. The scale bar represents 20 μm. Analyses were performed on three independent biological samples with three experimental replicates. F , schematic depiction of glycogen synthase activity regulation in vertebrates, which is downregulated by phosphorylation (including at Ser 641 ) via GSK3β (inhibited via insulin signaling), while glucose and glucose 6-phosphate upregulate GS activity in an allosteric fashion, and by PP1-mediated dephosphorylation (prompted by insulin signaling).

Article Snippet: Cells were kept for 1 h in blocking solution (1% bovine serum albumin in PBS), followed by a 16-h incubation at 4 to 8 °C with the respective primary antibody, anti-Glycogen Synthase (GS, total) (Cell signaling, #3893) or anti-Glycogen Synthase phosphorylated at Ser641 (pGS, inhibited) (Cell signaling, #3891), at 1:200 dilution in blocking solution.

Techniques: Expressing, Staining, Laser-Scanning Microscopy, Light Microscopy, Software, Activity Assay, Phospho-proteomics, De-Phosphorylation Assay

Journal: The Journal of Biological Chemistry

Article Title: Redox imbalance induces remodeling of glucose metabolism in Rhipicephalus microplus embryonic cell line

doi: 10.1016/j.jbc.2022.101599

Figure Lengend Snippet: The conserved axis of insulin pathway is closely correlated to glycogen metabolism and gluconeogenesis in BME26 cells. A , partial schematic depiction of the insulin signaling pathway and its contribution to glycogen metabolism. Indicated components were further analyzed. Straight arrows , single-step process; dotted arrow , multistep process; arrowhead , activation; blunt arrowhead , inhibition. B , BME26 cells respond to the addition of exogenous insulin (400 nM), recovering the viability lost by removing fetal bovine serum for 24 h ( C ) PI3K regulatory subunit (p85) gene silencing validation and its effects on glycogen synthase (GS) transcript levels ( D ) and glycogen content ( E ). Protein kinase B (AKT) and GSK3β gene silencing validation ( F and I , respectively), and its effects on GS ( G and J ) and PEPCK ( H and K ) transcript levels, respectively.

Article Snippet: Cells were kept for 1 h in blocking solution (1% bovine serum albumin in PBS), followed by a 16-h incubation at 4 to 8 °C with the respective primary antibody, anti-Glycogen Synthase (GS, total) (Cell signaling, #3893) or anti-Glycogen Synthase phosphorylated at Ser641 (pGS, inhibited) (Cell signaling, #3891), at 1:200 dilution in blocking solution.

Techniques: Activation Assay, Inhibition, Biomarker Discovery

Journal: The Journal of Biological Chemistry

Article Title: Redox imbalance induces remodeling of glucose metabolism in Rhipicephalus microplus embryonic cell line

doi: 10.1016/j.jbc.2022.101599

Figure Lengend Snippet: Metabolic remodeling involves simultaneous gluconeogenesis and mitochondrial activity in BME26 cells. A , viable NADP-ICDH knocked down BME26 cells after H 2 O 2 treatment. B , NADP-ICDH gene silencing validation and its effects on the transcript levels of glucose-6-phosphate dehydrogenase (G6PDH) ( C ), glycogen synthase kinase 3 β (GSK3β) ( D ), glycogen synthase (GS) ( E ) and phosphoenolpyruvate carboxykinase (PEPCK) ( F ), as measured 2 and 24 h after 2 or 4 mM H 2 O 2 addition, in BME26 cells. G , schematic representation of the proposed metabolic remodeling in BME26 in response to H 2 O 2 challenge. ∗ p <0.05; ∗∗ p <0.01; ∗∗∗∗ p < 0.0001; compared with the control without H 2 O 2 within each H 2 O 2 concentration of same dsRNA treatment; and # p <0.05; ## p <0.01; ### p <0.001; #### p <0.0001 compared with dsRNA nonrelated (dsGFP) within dsICDHnadp gene silencing, in Tukey’s multiple comparisons test.

Article Snippet: Cells were kept for 1 h in blocking solution (1% bovine serum albumin in PBS), followed by a 16-h incubation at 4 to 8 °C with the respective primary antibody, anti-Glycogen Synthase (GS, total) (Cell signaling, #3893) or anti-Glycogen Synthase phosphorylated at Ser641 (pGS, inhibited) (Cell signaling, #3891), at 1:200 dilution in blocking solution.

Techniques: Activity Assay, Biomarker Discovery, Control, Concentration Assay

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: Correlation of clinicopathological characteristics with UGP2 expression in the ZZU HCC cohort.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques: Expressing

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: UGP2 expression is frequently dysregulated in cancer. (a) UGP2 mRNA expression level from the TCGA dataset compared with normal tissues. (b) UGP2 protein expression in pancancer tissues and paired nontumour tissues. (c) Representative UGP2 histological scoring in pancancer tissues and paired nontumour tissues. UGP2: uridine diphosphate-glucose pyrophosphorylase 2; TCGA: The Cancer Genome Atlas.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques: Expressing

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: UGP2 mRNA levels and protein expression are significantly downregulated in HCC. (a) The UGP2 expression level was significantly lower in HCC tissues than in adjacent nontumour tissues in the TCGA cohort and GEO datasets. (b) A representative IHC image of UGP2 expression in HCC and normal tissues. (c) Representative images of UGP2 staining in HCC tissues. (d) The UGP2 expression level in HCC tissues was lower than that in paired nontumour tissues in the ZZU HCC cohort. UGP2: uridine diphosphate-glucose pyrophosphorylase 2; TCGA: The Cancer Genome Atlas; HCC: hepatocellular carcinoma; GEO: Gene Expression Omnibus; IHC: immunohistochemistry.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques: Expressing, Staining, Gene Expression, Immunohistochemistry

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: ROC curve analysis of UGP2 expression in HCC. (a–f) ROC curve analysis of UGP2 expression in HCC from the TCGA, GSE36376, GSE76297, GSE54236, GSE14520, and GSE64041 datasets. UGP2: uridine diphosphate-glucose pyrophosphorylase 2; TCGA: The Cancer Genome Atlas; HCC: hepatocellular carcinoma; ROC: receiver operating characteristic.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques: Expressing

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: Downregulated UGP2 expression predicts a poor prognosis in HCC patients in the TCGA cohort. (a, b) Kaplan-Meier survival curves showing the correlation between UGP2 expression levels and the OS/PFS of HCC patients in the TCGA cohort. (c, d) Kaplan-Meier analysis showed that OS/PFS was shorter in HCC patients in the TCGA cohort with low UGP2 expression levels regardless of the TNM stage. (e, f) GSEA of the relationship between low UGP2 expression levels and the survival of HCC patients in the TCGA cohort. UGP2: uridine diphosphate-glucose pyrophosphorylase 2; TCGA: The Cancer Genome Atlas; HCC: hepatocellular carcinoma; OS: overall survival; PFS: progression-free survival; HCC: hepatocellular carcinoma.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques: Expressing

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: Downregulated UGP2 expression predicts a poor prognosis in HCC patients in the ZZU cohort. (a) Kaplan-Meier survival curves showing the correlation between UGP2 expression levels and the OS of HCC patients in the ZZU cohort. (b) Kaplan-Meier analysis revealed that OS was short in HCC patients in the ZZU cohort with low UGP2 expression levels regardless of the TNM stage. UGP2: uridine diphosphate-glucose pyrophosphorylase 2; HCC: hepatocellular carcinoma; OS: overall survival; TNM: tumour-node-metastasis.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques: Expressing

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: Univariate and multivariate Cox regression analyses of risk factors for overall survival time in the ZZU HCC cohort.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques:

Journal: Disease Markers

Article Title: Low UGP2 Expression Is Associated with Tumour Progression and Predicts Poor Prognosis in Hepatocellular Carcinoma

doi: 10.1155/2020/3231273

Figure Lengend Snippet: UGP2 has potential roles in fatty acid metabolism. (a) KEGG functional enrichment analysis showing the potential mechanism of UGP2 in HCC. (b) GSEA of the relationship between low UGP2 expression and genes associated with fatty acid metabolism. (c–g) The scatter plots show that UGP2 expression is markedly correlated with genes involved in fatty acid metabolism. UGP2: uridine diphosphate-glucose pyrophosphorylase 2; KEGG: Kyoto Encyclopedia of Genes and Genomes; HCC: hepatocellular carcinoma.

Article Snippet: After 30 min of blocking with 10% bovine serum albumin, sections were incubated with a UGP2 primary antibody (1 : 100; ProteinTech Group, Inc.) at 4°C overnight.

Techniques: Functional Assay, Expressing

Journal: Brazilian Journal of Medical and Biological Research

Article Title: Induced maturation of hepatic progenitor cellsin vitro

doi: 10.1590/1414-431x20132455

Figure Lengend Snippet: Figure 3. Expression of liver cell markers during the induced maturation of hepatic progenitor cells (HPCs). Cells were treated with the combination condition of 2% HS+0.1 mM Dex+10 ng/mL HGF+20 ng/mL FGF4. A, The morphology of untreated and treated cells at D3, D6, D9, and D12 days after induction. Panels 1, 2, 3, and 4 are untreated cells; 7, 8, 9, and 10 are induced cells. Scale bar=200 mm. B, RT-PCR analysis of hepatic-related genes DLK, CK19, AFP, ALB, CK18, TAT, and ApoB at D0, D3, D6, D9, and D12 after induction. RT-PCR results were confirmed in at least three independent experiments, and representative results are shown. C, Immunofluorescence staining of DLK, AFP, ALB, and UGT1A markers at D0, D3, D6, D9, and D12 days after induction. Negative controls (NC) are cells stained with nonspecific IgG. Scale bar=200 mm. D, Real-time PCR analysis of hepatic related genes AFP, ALB, CK18, TAT of the induced cells at D12 compared with untreated cells as negative controls and adult liver cells as positive controls. See Figure 1 for explanation of abbreviations. *P,0.05 induced D12 vs control D12 (two-tailed Student t-test); **P,0.05 LC14d vs induced D12 (two-tailed Student t-test).

Article Snippet: As previously described (13), methanol was used to fix the treated cells at -206C for 15 min, 5% goat serum was used to block cells at room temperature (RT) for 1 h. Then, cells were incubated with primary antibodies against delta-like protein (DLK), ALB, a-fetoprotein (AFP), or glucuronosyltransferase 1 A (UGT1A) (Santa Cruz Biotechnology, USA) at RT for 1 h, followed by incubation with DyLight 488-labeled secondary antibodies (Jackson ImmunoResearch Laboratories, USA) at RT for 30 min.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Control, Two Tailed Test

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Levels of glycogen metabolizing enzymes in uterine homogenates. (a–d) Western blots for hexokinase 1 (HK1, a), phospho‐glycogen synthase (pGYS, b), glycogen synthase (GYS, c), and glycogen phosphorylase (PYG, d) in uteri collected from mice at proestrus (PROE) and days postcoitum (DPC) 1.5, 3.5, and 5.5. n = 4.

Article Snippet: Glycogen synthase , 3886 Cell signaling , WB , 1:500 , BSA.

Techniques: Western Blot

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Localization of glycogen synthesizing enzymes in the endometrium from proestrus (PROE) until days postcoitum (DPC) 5.5. Immunohistochemistry for hexokinase 1 (HK1) and glycogen synthase (GYS) in uteri collected at PROE, DPC 1.5, DPC 3.5, and DPC 5.5 IIS. n = 4. Scale bar = 50 µm. IIS, interimplantation site; Neg, negative control.

Article Snippet: Glycogen synthase , 3886 Cell signaling , WB , 1:500 , BSA.

Techniques: Immunohistochemistry, Negative Control

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Immunostaining for glycogen metabolizing enzymes at the implantation site (IS) and the interimplantation site (IIS) on DPC 5.5. Immunohistochemistry for hexokinase 1 (HK1), glycogen synthase (GYS), glycogen phosphorylase (PYG), and glucose‐6‐phosphatase (G6PC) in uteri at DPC 5.5. n = 4. Scale bar = 50 µm. DPC, days postcoitum; Neg, negative control.

Article Snippet: Glycogen synthase , 3886 Cell signaling , WB , 1:500 , BSA.

Techniques: Immunostaining, Immunohistochemistry, Negative Control

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Western blots comparing levels of glycogen metabolizing enzymes in the uterus at the interimplantation site (IIS) and implantation site (IS) on DPC 5.5. Levels of hexokinase 1 (HK1, a), phospho‐glycogen synthase (pGYS, b), glycogen synthase (GYS, c), and glycogen phosphorylase (PYG, d). * p < 0.05 relative to DPC 5.5‐IIS. n = 4–5. DPC, days postcoitum.

Article Snippet: Glycogen synthase , 3886 Cell signaling , WB , 1:500 , BSA.

Techniques: Western Blot

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Localization of glycogen metabolizing enzymes in the decidualized and undecidualized uterine horn in an artificial decidualization model. Immunohistochemistry for hexokinase 1 (HK1), glycogen synthase (GYS), glycogen phosphorylase (PYG), and glucose‐6‐phosphatase (G6PC) in hormonally primed mice. One horn was simulated to decidualize. The unstimulated horn served as a nondecidualized control. n = 4. Scale bar = 50 µm.

Article Snippet: Glycogen synthase , 3886 Cell signaling , WB , 1:500 , BSA.

Techniques: Immunohistochemistry, Control

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Levels of glycogen metabolizing enzymes in uterine horns stimulated to decidualize or left unstimulated. (a–d) Western blots for hexokinase 1 (HK1, a), phospho‐glycogen synthase (pGYS, b), glycogen synthase (GYS, c), and glycogen phosphorylase (PYG, d) in uterine horns stimulated to decidualization with corn oil or unstimulated after hormonal priming. ** p < 0.01 relative to unstimulated horn. n = 5.

Article Snippet: Glycogen synthase , 3886 Cell signaling , WB , 1:500 , BSA.

Techniques: Western Blot

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Primary antibodies and summary of conditions used for western blot (WB) and immunohistochemistry (IHC)

Article Snippet: Glycogen synthase , 3886 Cell signaling , WB , 1:500 , BSA.

Techniques: Western Blot, Immunohistochemistry, Blocking Assay

Journal: Molecular Oncology

Article Title: Consensus molecular subtyping of colorectal carcinoma brain metastases reveals a metabolic signature associated with poor patient survival

doi: 10.1002/1878-0261.13748

Figure Lengend Snippet: The identified metabolic gene signature is superior to consensus molecular subtype (CMS) in predicting survival of colorectal cancer (CRC) patients with brain metastasis. Overall survival (OS) of CRC patients suffering from liver or brain metastases stratified according to the CMS classification of the metastatic tissue. Survival impacts were compared between (A) all four subtypes (CMS1 vs. CMS2 vs. CMS3 vs. CMS4), (B) single subtypes and all other subtypes (CMS4 vs. others for CRC liver, CMS3 vs. others for CRC brain), and (C) high and low expression of the seven metabolic signature genes CHAC2 , CA8 , PIK3R3 , CYP26B1 , DHRS9 , UGT8 , and RET within brain metastases. An optimal cut‐off value was calculated to define high/low groups based on the transcriptomic data from metastatic samples. Survival data is shown as Kaplan–Meier curves and differences were tested with a log‐rank test. Hazard ratios with 95% confidence interval and corresponding P ‐values are indicated for all comparisons between two cohorts.

Article Snippet: Shortly, after deparaffinization and peroxidase activity blocking with 5% H 2 O 2 , the samples were exposed to the following primary antibodies: DHRS9 (#14560‐1‐AP), CA8 (#12391‐1‐AP), CYP26B1 (#21555‐1‐AP), PIK3R3 (#27035‐1‐AP), UGT8 (#17982‐1‐AP, all from Proteintech, Rosemont, IL, USA), CHAC2 (#HPA049235; Sigma‐Aldrich, St. Louis, MO, USA) and RET (#ab134100; Abcam, Cambridge, UK).

Techniques: Expressing

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Levels of glycogen metabolizing enzymes in uterine homogenates. (a–d) Western blots for hexokinase 1 (HK1, a), phospho‐glycogen synthase (pGYS, b), glycogen synthase (GYS, c), and glycogen phosphorylase (PYG, d) in uteri collected from mice at proestrus (PROE) and days postcoitum (DPC) 1.5, 3.5, and 5.5. n = 4.

Article Snippet: Phospho‐glycogen synthase , 47043 Cell signaling , WB , 1:500 , Milk.

Techniques: Western Blot

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Localization of glycogen synthesizing enzymes in the endometrium from proestrus (PROE) until days postcoitum (DPC) 5.5. Immunohistochemistry for hexokinase 1 (HK1) and glycogen synthase (GYS) in uteri collected at PROE, DPC 1.5, DPC 3.5, and DPC 5.5 IIS. n = 4. Scale bar = 50 µm. IIS, interimplantation site; Neg, negative control.

Article Snippet: Phospho‐glycogen synthase , 47043 Cell signaling , WB , 1:500 , Milk.

Techniques: Immunohistochemistry, Negative Control

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Immunostaining for glycogen metabolizing enzymes at the implantation site (IS) and the interimplantation site (IIS) on DPC 5.5. Immunohistochemistry for hexokinase 1 (HK1), glycogen synthase (GYS), glycogen phosphorylase (PYG), and glucose‐6‐phosphatase (G6PC) in uteri at DPC 5.5. n = 4. Scale bar = 50 µm. DPC, days postcoitum; Neg, negative control.

Article Snippet: Phospho‐glycogen synthase , 47043 Cell signaling , WB , 1:500 , Milk.

Techniques: Immunostaining, Immunohistochemistry, Negative Control

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Western blots comparing levels of glycogen metabolizing enzymes in the uterus at the interimplantation site (IIS) and implantation site (IS) on DPC 5.5. Levels of hexokinase 1 (HK1, a), phospho‐glycogen synthase (pGYS, b), glycogen synthase (GYS, c), and glycogen phosphorylase (PYG, d). * p < 0.05 relative to DPC 5.5‐IIS. n = 4–5. DPC, days postcoitum.

Article Snippet: Phospho‐glycogen synthase , 47043 Cell signaling , WB , 1:500 , Milk.

Techniques: Western Blot

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Localization of glycogen metabolizing enzymes in the decidualized and undecidualized uterine horn in an artificial decidualization model. Immunohistochemistry for hexokinase 1 (HK1), glycogen synthase (GYS), glycogen phosphorylase (PYG), and glucose‐6‐phosphatase (G6PC) in hormonally primed mice. One horn was simulated to decidualize. The unstimulated horn served as a nondecidualized control. n = 4. Scale bar = 50 µm.

Article Snippet: Phospho‐glycogen synthase , 47043 Cell signaling , WB , 1:500 , Milk.

Techniques: Immunohistochemistry, Control

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Levels of glycogen metabolizing enzymes in uterine horns stimulated to decidualize or left unstimulated. (a–d) Western blots for hexokinase 1 (HK1, a), phospho‐glycogen synthase (pGYS, b), glycogen synthase (GYS, c), and glycogen phosphorylase (PYG, d) in uterine horns stimulated to decidualization with corn oil or unstimulated after hormonal priming. ** p < 0.01 relative to unstimulated horn. n = 5.

Article Snippet: Phospho‐glycogen synthase , 47043 Cell signaling , WB , 1:500 , Milk.

Techniques: Western Blot

Journal: Molecular Reproduction and Development

Article Title: Endometrial glycogen metabolism during early pregnancy in mice

doi: 10.1002/mrd.23634

Figure Lengend Snippet: Primary antibodies and summary of conditions used for western blot (WB) and immunohistochemistry (IHC)

Article Snippet: Phospho‐glycogen synthase , 47043 Cell signaling , WB , 1:500 , Milk.

Techniques: Western Blot, Immunohistochemistry, Blocking Assay