Journal: Molecular & Cellular Proteomics : MCP

Article Title: Integrated Systems Analysis of the Murine and Human Pancreatic Cancer Glycomes Reveals a Tumor-Promoting Role for ST6GAL1

doi: 10.1016/j.mcpro.2021.100160

Figure Lengend Snippet: Transcriptomic analysis identifies ST6GAL1 and ST3GAL3 as enriched in the cancerous ducts. A , biosynthetic pathways for glycans underlying select lectin signatures are shown. ST3GALs are responsible for transferring α-2,3-sialosides and ST6GAL1/2 for α-2,6-sialosides, and MGAT3 for bisecting GlcNAc. The glycans are annotated following the Symbolic Nomenclature for Glycans. B , transcriptomic analysis assessing the mRNA levels of select glycosyltransferases between normal adjacent and matched cancerous tissues within the same patient. C , uMAP plots representing the cells isolated from patients with PDAC (n = 24) and normal pancreata (n = 11) pooled on single-cell sequencing. The clusters representing the normal ductal cells and tumor ductal cells are highlighted. D , violin plot showing the comparison of ST6Gal1 levels in normal ( green ) versus PDAC ( blue ) ductal clusters. The clusters in each group are combined. The violin plots showing the comparison of ST6Gal1 and ST3GAL1 levels in normal ( green ) versus PDAC ( blue ) ductal clusters. The clusters in each group are combined. ns, p > 0.05; ∗ p < 0.05; student’s t test ( two-tailed ). MGAT3, beta-1,4-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase; ns, not statistical; PDAC, pancreatic ductal adenocarcinoma; ST3GAL3, ST3 beta-galactoside alpha-2,3-sialyltransferase 3; ST6GAL1, ST6 beta-galactoside alpha-2,6-sialyltransferase 1; uMAP, Uniform Manifold Approximation and Projection.

Article Snippet: The slide was then incubated with anti-human ST6GAL1 (Proteintech, cat # 14355-1-AP) followed by Opal Polymer horseradish peroxidase Ms + Rb (Akoya Biosciences, cat # ARH1001EA) and tyramide-linked 650 Opal fluorophore (Akoya Biosciences, cat # FP1496001KT) to amplify signals.

Techniques: Transferring, Isolation, Sequencing, Comparison, Two Tailed Test

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Integrated Systems Analysis of the Murine and Human Pancreatic Cancer Glycomes Reveals a Tumor-Promoting Role for ST6GAL1

doi: 10.1016/j.mcpro.2021.100160

Figure Lengend Snippet: Profiling of ST6GAL1 and SNA in human pancreatic cancer shows association with the stage and survival. A , H&E of the normal pancreas ( left ), stage I pancreatic adenocarcinoma ( center ), and stage IV PDAC ( right ) stained from a BioMax human tissue microarray. B , multiplex OPAL IF staining of SNA ( yellow ), ST6GAL1 ( red ), and DAPI ( blue ) on the corresponding normal pancreas and stage I and stage IV pancreatic adenocarcinoma purchased from BioMax human tissue microarray. The scale bars represent 25 μm. C , quantification of SNA-positive cells per high-powered field based on multiplex IF in normal versus all cancerous cases in human tissue microarray. Owing to the high number of positive cells, each HPF was given a score (1–3) based on the number of SNA-positive cells per field. D , quantification of ST6GAL1-positive cells per high-powered field in normal versus all cancerous cases in human tissue microarray. E , distribution of the total ST6GAL1-positive cells per high-powered field across normal and each stage of pancreatic cancer based on multiplex IF imaging of human pancreatic cancer tissue microarray. Statistical significance was evaluated using the Student’s t test ( two-tailed ): ns, p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. HFP, high-powered field; ns, not statistical; PDAC, pancreatic ductal adenocarcinoma; SNA, Sambucus nigra agglutinin; ST6GAL1, ST6 beta-galactoside alpha-2,6-sialyltransferase 1.

Article Snippet: The slide was then incubated with anti-human ST6GAL1 (Proteintech, cat # 14355-1-AP) followed by Opal Polymer horseradish peroxidase Ms + Rb (Akoya Biosciences, cat # ARH1001EA) and tyramide-linked 650 Opal fluorophore (Akoya Biosciences, cat # FP1496001KT) to amplify signals.

Techniques: Staining, Microarray, Multiplex Assay, Imaging, Two Tailed Test

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Integrated Systems Analysis of the Murine and Human Pancreatic Cancer Glycomes Reveals a Tumor-Promoting Role for ST6GAL1

doi: 10.1016/j.mcpro.2021.100160

Figure Lengend Snippet: Pancreas-specific deletion of ST6GAL1 reduces disease burden in murine PDAC. A , breeding schematic illustrating the generation of novel ST6KC mice. The offspring of parental strains crossed into p48-Cre mice drive the induction of mutant KRAS G12D and deletion of ST6GAL1 under the same promoter. B , IHC staining for ST6GAL1 ( brown ) in KC and ST6KC mice (n = 3 per group). The number of ST6GAL1+ cells per high-powered field is quantified. The scale bar represents 50 μm. C , IF staining for SNA ( yellow ) and DAPI ( blue ) in KC and ST6KC mice (n = 3 per group). The number of SNA+ cells per high-powered field is quantified. The scale bar represents 100 μm. D , H&E of 14-week-old FFPE pancreata from KC and ST6KC mice (n = 5 per group). The percent of the preserved normal pancreas area is quantified per high-powered field. The scale bar represents 200 μm. E , trichrome and gomori ( blue ) stain of FFPE pancreata from 14-week-old KC and ST6KC mice (n = 5 per group). The percent of collagen deposition fibrosis is quantified per high-powered field on the right. The scale bar represents 200 μm. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; Student’s t test ( two-tailed ). FFPE, formalin-fixed paraffin-embedded; IF, immunofluorescence; IHC, immunohistochemical; PDAC, pancreatic ductal adenocarcinoma; SNA, Sambucus nigra agglutinin; ST6GAL1, ST6 beta-galactoside alpha-2,6-sialyltransferase 1; ST6KC, ST6GAL1 flx/flx ;p48 Cre ; LSL KRASG12D .

Article Snippet: The slide was then incubated with anti-human ST6GAL1 (Proteintech, cat # 14355-1-AP) followed by Opal Polymer horseradish peroxidase Ms + Rb (Akoya Biosciences, cat # ARH1001EA) and tyramide-linked 650 Opal fluorophore (Akoya Biosciences, cat # FP1496001KT) to amplify signals.

Techniques: Mutagenesis, Immunohistochemistry, Staining, Two Tailed Test, Formalin-fixed Paraffin-Embedded, Immunofluorescence, Immunohistochemical staining

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: ST6GAL1-mediated sialylation inhibits the antitumor immune response in colorectal cancer

doi: 10.1007/s13402-025-01118-w

Figure Lengend Snippet: ST6GAL1 expression is substantially increased in patients with colorectal cancer (CRC) resistant to neoadjuvant therapy. ST6GAL1 expression was measured in 14 surgically removed fresh CRC tumor tissues and matched adjacent normal tissues using RT‒qPCR, WB and IHC after. The correlation between ST6GAL1 expression and sensitivity to neoadjuvant therapy was analyzed in combination with the clinical information. ( A ) ST6GAL1 mRNA expression was quantified by RT‒qPCR in tumor tissues compared with adjacent normal tissues from 14 clinical CRC patients. ( B ) Statistical analysis of the S T6GAL1 mRNA level in the CRC patients in A. ( C ) ST6GAL1 protein levels were measured in 14 clinical samples from CRC patients by WB. ( D ) Statistical analysis of the ST6GAL1 protein levels in CRC patients in C. ( E ) Representative images of HE staining and IHC staining of ST6GAL1 in colorectal tumor tissue and adjacent normal colorectal tissue from CRC patients (scale bars, 625 μm; scale bars, 50 μm). ( F ) Statistical results for the relative intensity of ST6GAL1 staining shown in E. ( G ) Statistical analysis of the correlation between ST6GAL1 mRNA expression and sensitivity to neoadjuvant therapy. ( H ) Representative images of IHC staining for ST6AGL1 in colorectal tumor tissue from neoadjuvant therapy-sensitive and nonsensitive CRC patients (scale bars, 625 μm; scale bars, 50 μm). ( I ) Statistical results for the relative intensity of ST6GAL1 staining shown in H. Statistical significance was determined using Student’s t test ( B , D , F and G )

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against ST6GAL1 (AF5924; R&D Systems) and visualized using the Dako REALTM EnVisionTM detection system (Dako, K5007).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: ST6GAL1-mediated sialylation inhibits the antitumor immune response in colorectal cancer

doi: 10.1007/s13402-025-01118-w

Figure Lengend Snippet: ST6GAL1 expression is negatively correlated with the immune response in CRC. scRNA-seq data from public CRC databases were used to analyze ST6GAL1 expression in different cell subsets and its correlation with immune response pathways. ( A ) Uniform manifold approximation and projection (UMAP) plots showing the clusters of cells in the tumor tissue. ( B ) ST6GAL1 expression in different cell clusters. ( C ) GSEA of immune response-associated pathways between ST6GAL1 + cells and ST6GAL1 − cells in tumor tissues. ST6GAL1 expression and IFNG sensitivity in different CRC cell lines were further verified by RT‒qPCR and WB. ( D ) ST6GAL1 mRNA expression in the different CRC cell lines was quantified by RT‒qPCR. ( E ) ST6GAL1 protein levels were measured in 4 CRC cell lines using WB. ( F ) The mRNA levels of molecules associated with the IFNG response were measured by RT‒qPCR in different CRC cell lines treated with 100 ng/µl IFNG for 24 h

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against ST6GAL1 (AF5924; R&D Systems) and visualized using the Dako REALTM EnVisionTM detection system (Dako, K5007).

Techniques: Expressing

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: ST6GAL1-mediated sialylation inhibits the antitumor immune response in colorectal cancer

doi: 10.1007/s13402-025-01118-w

Figure Lengend Snippet: ST6GAL1 expression decreases IFNG sensitivity in CRC cell lines. After ST6GAL1 overexpression (OE) in the SW48 cell line (SW48-OE) and the HT29 cell line (HT29-OE) and ST6GAL1 knockdown (KD) in the Caco2 cell line (Caco2-KD) and the HT29 cell line (HT29-KD), as well as the construction of the related empty virus (EV) control cell lines SW48-EV, Caco2-KD and HT29-EV, the levels of molecules associated with the IFNG response were measured. ( A , B , C and D ) The mRNA levels of IFNG response-associated molecules were measured by RT‒qPCR in the SW48-EV and SW48-OE cell lines ( A ), the HT29-EV and HT29-OE cell lines ( B ), the Caco2-EV and Caco2-KD cell lines ( C ), and the HT29-EV and HT29-KD cell lines ( D ) after treatment with 100 ng/µl IFNG for 24 h. ( E , F , and G ) GBP1 and IRF1 protein levels in the SW48-EV and SW48-OE cell lines ( E ), the Caco2-EV and Caco2-KD cell lines ( F ), and the HT29-EV, HT29-OE and HT29-KD cell lines ( G ) were measured by WB after treatment with 100 ng/µl IFNG for 12 and 24 h

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against ST6GAL1 (AF5924; R&D Systems) and visualized using the Dako REALTM EnVisionTM detection system (Dako, K5007).

Techniques: Expressing, Over Expression, Knockdown, Virus, Control

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: ST6GAL1-mediated sialylation inhibits the antitumor immune response in colorectal cancer

doi: 10.1007/s13402-025-01118-w

Figure Lengend Snippet: ST6GAL1 overexpression promotes the immune escape of CRC tumor cells in vivo. An ST6GAL1 -OE MC38 cell line (MC38-OE) and the related empty virus (EV)-expressing control cell line MC38-EV were constructed. We used MC38-EV ( n = 4) and MC38-OE ( n = 4) cells to construct an animal model of subcutaneous tumorigenesis in wild-type C57BL/6 mice. When the tumor grew to an appropriate size, the tumor was removed, and the change in cell count was detected by flow cytometry. ( A ) Statistical analysis of the differences in the numbers of cells in tumors, draining lymph nodes and PBMCs between the ST6GAL1-EV and ST6GAL1-OE groups. ( B ) Flow cytometry analyses of CD3 + T, CD4 + T, CD8 + T and Treg cells in CD45-gated cells from tumor tissues from the ST6GAL1-EV and ST6GAL1-OE groups. ( C , D and E ) Statistical results for CD3 + T-cell numbers, CD4 + and CD8 + T-cell numbers ( D ) and Treg numbers ( E ) between the ST6GAL1-EV and ST6GAL1-OE groups. ( F , H and J ) Flow cytometry analyses of GZMB from CD8 + T cells ( F ), IFNG from CD4 + T cells ( H ) and IFNG from CD8 + T cells ( J ) in tumor tissues from the ST6GAL1-EV and ST6GAL1-OE groups. ( G , I and K ) Statistical results for GZMB from CD8 + T cells ( G ), IFNG from CD4 + T cells ( I ) and IFNG from CD8 + T cells ( K ) in tumor tissues from the ST6GAL1-EV and ST6GAL1-OE groups. Statistical significance was determined using Student’s t test ( A , C , D , E , G , I and K )

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against ST6GAL1 (AF5924; R&D Systems) and visualized using the Dako REALTM EnVisionTM detection system (Dako, K5007).

Techniques: Over Expression, In Vivo, Virus, Expressing, Control, Construct, Animal Model, Cell Counting, Flow Cytometry

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: ST6GAL1-mediated sialylation inhibits the antitumor immune response in colorectal cancer

doi: 10.1007/s13402-025-01118-w

Figure Lengend Snippet: Multiomics analysis revealed that ST6GAL1 inhibits the activity of the IFNG signaling pathway. SW48-EV ( n = 3) and SW48-OE ( n = 3) cell lines were subjected to RNA-sequencing (RNA-seq) after treatment with 100 ng/µl IFNG for 24 h ( A , B and C ). ( A ) Volcano plot showing all genes expressed in the SW48-EV and SW48-OE cell lines. The x-axis shows the log 2 fold change (SW48-OE vs. SW48-EV cells), and the y-axis shows the -log 10 P value, which represents the threshold values in log transformation. Each dot represents a differentially expressed gene (DEG). The red dots indicate significantly upregulated DEGs, the blue dots indicate significantly downregulated DEGs, and the gray dots represent DEGs whose differential expression was not statistically significant. ( B ) Heatmap showing the relative expression of selected genes in three different functional modules (immune-related genes, inflammatory factor-related genes and IFNG response-related genes) in the S6TGAL1-OE cell line. ( C ) Gene set enrichment analysis (GSEA) of the SW48-OE cell line revealed an upregulated signaling pathway compared with that of the SW48-EV cell line. Kinase phosphorylation array detection of the SW48-EV and SW48-OE cell lines was subsequently performed ( D and E ). ( D ) Results of the IFNGR1-pTyr457 measurement via kinase phosphorylation array in the SW48-EV and SW48-OE cell lines. ( E ) KEGG pathway analysis of the differentially phosphorylated proteins in SW48-EV and SW48-OE cell lines identified via kinase phosphorylation array detection

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against ST6GAL1 (AF5924; R&D Systems) and visualized using the Dako REALTM EnVisionTM detection system (Dako, K5007).

Techniques: Activity Assay, RNA Sequencing, Transformation Assay, Quantitative Proteomics, Expressing, Functional Assay, Phospho-proteomics

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: ST6GAL1-mediated sialylation inhibits the antitumor immune response in colorectal cancer

doi: 10.1007/s13402-025-01118-w

Figure Lengend Snippet: ST6GAL1 regulates the IFNGR1/JAK1/STAT1 signaling pathway by affecting IFNGR1 and BICD2 binding. ( A , C and E ) WBs showing the levels of the pIFNGR1, IFNGR1, and pSTAT1 proteins in the SW48-EV and SW48-OE cell lines ( A ), the Caco2-EV and Caco2-KD cell lines ( C ), and the HT29-EV, HT29-OE and HT29-KD cell lines ( E ) after treatment with 100 ng/µl IFNG for 15 and 30 min ( B , D , and F ) Statistical results for the gray values shown in A ( B ), C ( D ) and E ( F ). ( G ) IP results for the HT29-EV and HT29-OE cell lines. ( H ) Immunofluorescence staining for SNA-FITC (green) and BICD2 (red) in the Caco2-EV and Caco2-KD cell lines. Nuclei were counterstained with DAPI (blue). Scale bars, 20 μm

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against ST6GAL1 (AF5924; R&D Systems) and visualized using the Dako REALTM EnVisionTM detection system (Dako, K5007).

Techniques: Binding Assay, Immunofluorescence, Staining

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: ST6GAL1-mediated sialylation inhibits the antitumor immune response in colorectal cancer

doi: 10.1007/s13402-025-01118-w

Figure Lengend Snippet: The mechanism by which ST6GAL1 promotes tumor immune evasion. ST6GAL1, an enzyme that adds sialic acid to proteins, is abundantly localized in the tumor microenvironment, where it inhibits IFNGR1 phosphorylation through the sialylation of BICD2, thereby suppressing the activation of the IFNGR1/JAK1/STAT1 signaling pathway. This interaction leads to resistance to the antitumor immune response and CRC development. *BICD2, BICD cargo adaptor 2; GBP1, guanylate binding protein 1; IDO1, indoleamine 2,3-dioxygenase 1; ISGs, IFN-stimulating genes; IRF, interferon regulatory factor; IFNG, interferon gamma; IFNGR, interferon gamma receptor; JAK, Janus kinase; NK, natural killer; STAT1, signal transducer and activator of transcription 1; ST6GAL1, ST6 β-galactoside α2,6-sialyltransferase; TH1, helper T

Article Snippet: Paraffin-embedded tissue sections were stained with antibodies against ST6GAL1 (AF5924; R&D Systems) and visualized using the Dako REALTM EnVisionTM detection system (Dako, K5007).

Techniques: Phospho-proteomics, Activation Assay, Binding Assay

Journal: Redox Biology

Article Title: A Golgi-associated redox switch regulates catalytic activation and cooperative functioning of ST6Gal-I with B4GalT-I

doi: 10.1016/j.redox.2019.101182

Figure Lengend Snippet: Functional consequences of the inactivation of the ST6Gal-I. a) Immunoblotting of the ST6Gal and its ST6Gal-I CysDM variant in COS-7 cells. Cell transfections were performed by electroporation before SDS-PAGE and immunoblotting with ST6Gal-specific antibodies (anti-CD75) as above. b) Representative figures on SNA lectin binding to the cells transfected separately with the same plasmids. Transfections and lectin staining was carried out as describe above. c) Quantification of the SNA lectin binding to cells using the high content imaging system. The data are shown as relative changes (mean % + SD, n = 3, 70–90000 cells/assay). d) Live cell proliferation rate analysis of COS-7 cells transfected with an empty vector (EV) or with vectors expressing the wild-type ST6Gal-I (WT) or the cysteine double mutant of ST6Gal-I (CysDM). Data are expressed as mean confluence % (±SD (n = 5) using a build-in Incyte™ protocol. The star (*) mark the statistically significant difference (p < 0.05) between relative confluence values of the wild-type ST6Gal-I (WT) and the cysteine double mutant (ST6Gal-I CysDM ) at each time point. e) Live cell wound closure analysis of wild type (WT) and double cysteine (CysDM) ST6Gal-I variant expressing cells. The graph shows thee mean confluence (% ±SD (n = 4) at the indicated times after making the wound. Significant changes between confluence values between the wild-type ST6Gal-I (WT) and cysteine double mutant of ST6Gal-I (CysDM) at each time point are marked by stars (p < 0.05*). f) Representative images of the wound closure in COS-7 cells transfected with an empty vector (EV), ST6Gal-I (WT) or the cysteine double mutant of ST6Gal-I (CysDM) at 0 h, 60 h and 120 h time point after introduction of the wound. Scale bar 300 μm. Green color represents the original wound, and yellow color depicts the area of repopulated cells. g) Sensitivity cells to staurosporine-induced apoptosis. Cells transfected with the indicated ST6Gal-I variants were subjected to 5 μM staurosporine for 0–6 h. After fixing, cells were stained with the Hoechst dye before imaging high content imaging system (Operetta, PerkinElmer). Fragmentation index of nuclei was then calculated after image segmentation using the “RMS Nuclear Fragmentation” built-in protocol, in which the coefficient of variance percentage (CV%) of the nuclear intensity is used to calculate the fragmentation index. Healthy nuclei have low CV values whereas fragmented nuclei have higher CV values. The values represent the mean (±SD, n = 3, 15,000 cells each. Statistically significant changes are marked with stars (p < 0.05*, p < 0.01**, p < 0.001***). h-k) The effect of the wild type ST6Gal-I and its double cysteine variant on nuclear and cell morphology. Cells transfected with the indicated constructs were plated on a 96 well plate, fixed immuno-stained with an α-tubulin antibody, and counterstained with the Hoechst dye. After imaging (Operetta, PerkinElmer) and segmentation, in-built “Harmony” protocols were used to calculate nuclear and cytoplasmic areas (h and j) and their roundness (i and k). The data are expressed as the mean % (±SD, n = 3, 15,000 cells each) of the control values (EV = empty vector). No statistically significant changes were detected between the different ST6Gal-I transfectants (p > 0.05).

Article Snippet: For normalization, identical cell samples were subjected to SDS-PAGE and western blotting with the anti-CD75 antibody (#AP21150PU-N, Acris Antibodies GmbH, Germany) to determine the level of ST6Gal-I protein in each sample (Suppl.).

Techniques: Functional Assay, Western Blot, Variant Assay, Transfection, Electroporation, SDS Page, Binding Assay, Staining, Imaging, Plasmid Preparation, Expressing, Mutagenesis, Construct, Control

Journal: Gut microbes

Article Title: Sialic acid-based probiotic intervention in lactating mothers improves the neonatal gut microbiota and immune responses by regulating sialylated milk oligosaccharide synthesis via the gut-breast axis.

doi: 10.1080/19490976.2024.2334967

Figure Lengend Snippet: Figure 2. SA+Pro intervention in lactating St6gal1± (H) rats compromised allergic responses in neonates by regulating 6′-SL synthesis and the neonatal gut microbiota. (a) A schematic diagram depicting the cross-feeding procedure used with WT neonates fed by WT, H, and KO maternal rats. (b) Comparison of the major milk oligosaccharides in different rat mothers. (c) Changes in the intestinal microbiota of neonates fed by different maternal rats. (d) The serum levels of cytokines in neonatal rats and changes in lymphocyte subsets in the spleen and neonatal MLNs. (e) A schematic diagram indicating that neonates fed by H maternal rats or H maternal rats received SA+Pro intervention. (f) Changes in the major milk oligosaccharides of the H maternal rats, with or without SA+Pro intervention. (g) The abundance of intestinal bacterial genera and the total and individual SCFA contents in neonatal rats. (h) A schematic diagram representing the establishment of a model of OVA-induced allergic responses in neonates fed by H maternal rats,

Article Snippet: Antibodies against GPR41 (66811–1-Ig, Sanying, WuHan, China) PI3K (A4992, ABclonal, Wuhan, China), phosphorylated (p)PI3K (Ab278545, Abcam, UK), Akt (A18675, ABclonal, Wuhan, China), p-Akt (AP1208, ABclonal, Wuhan, China), mTOR (A11355, ABclonal, Wuhan, China), p-mTOR (AP0115, ABclonal, Wuhan, China), RxRα (A19105, ABclonal, Wuhan, China), SREBP1 (A5754, ABclonal, Wuhan, China), PPARγ (A16958, ABclonal, Wuhan, China), St6gal1 (A5754, ABclonal, Wuhan, China), and GAPDH (ABP-R001, Xianzhi, Hangzhou, China) were used as primary antibodies.

Techniques: Comparison

Journal: Gut microbes

Article Title: Sialic acid-based probiotic intervention in lactating mothers improves the neonatal gut microbiota and immune responses by regulating sialylated milk oligosaccharide synthesis via the gut-breast axis.

doi: 10.1080/19490976.2024.2334967

Figure Lengend Snippet: Figure 3. The Gpr41-PI3K-Akt-PPAR pathway was involved in regulating 6′-SL biosynthesis in mammary glands by SA+Pro. (a) Bubble plots representing KEGG enrichment analysis of differentially expressed genes related to signaling pathways in the mammary glands of different groups. The CON group indicated the mammary glands of the WT maternal mice, the H groups indicated the mammary glands of the H maternal mice, and the SA and SA+Pro groups indicated the mammary glands of the maternal mice that received SA and SA+Pro intervention during lactation, respectively. (b) Bubble plots represent GO enrichment analysis of differentially expressed genes in the mammary glands of different groups. (c) Expression of glycosyltransferases in the mammary glands of rats in different groups. (d) Effects of different treatments on the expression of glycosyltransferases in human mammary epithelial cells (MCF-10A). (e) The expression and phosphorylation levels of proteins involved in the PI3K-AKT-mTOR and PPARγ pathways were determined using western blot analysis. GAPDH expression was detected as an internal control. The protein expression levels were quantified using densitometry. (f) Effects of PPARγ and PI3K-AKT inhibitors on ST6Gal-1 expression in MCF-10A cells stimulated with But. (g) ChIP assay results depicting the interaction between the RXRα–PPARγ complex and the promoter region of St6gal1. All values are presented as the mean ± SD (n ≥ 3), ***p < .001.

Article Snippet: Antibodies against GPR41 (66811–1-Ig, Sanying, WuHan, China) PI3K (A4992, ABclonal, Wuhan, China), phosphorylated (p)PI3K (Ab278545, Abcam, UK), Akt (A18675, ABclonal, Wuhan, China), p-Akt (AP1208, ABclonal, Wuhan, China), mTOR (A11355, ABclonal, Wuhan, China), p-mTOR (AP0115, ABclonal, Wuhan, China), RxRα (A19105, ABclonal, Wuhan, China), SREBP1 (A5754, ABclonal, Wuhan, China), PPARγ (A16958, ABclonal, Wuhan, China), St6gal1 (A5754, ABclonal, Wuhan, China), and GAPDH (ABP-R001, Xianzhi, Hangzhou, China) were used as primary antibodies.

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Western Blot, Control

Journal: PLoS ONE

Article Title: An Enhanced Monomeric Blue Fluorescent Protein with the High Chemical Stability of the Chromophore

doi: 10.1371/journal.pone.0028674

Figure Lengend Snippet: (A–M) C-terminal fusion constructs . For each fusion protein, the linker amino acid length is indicated after the name of the targeted organelle or fusion protein. (A) mTagBFP2-lamin B1-10; (B) mTagBFP2-CAAX-farnesyl-5; (C) mTagBFP2-endoplasmic reticulum-5 (calreticulin and KDEL); (D) mTagBFP2-fibrillarin-7; (E) mTagBFP2-light chain clathrin-15; (F) mTagBFP2-β-actin-7; (G) mTagBFP2-caveolin 1-10; (H) mTagBFP2-vinculin-22; (I) mTagBFP2-CAF1-10 (chromatin assembly factor); (J) mTagBFP2-Rab5a-7; (K) mTagBFP2-α-tubulin-18; (L) mTagBFP2-myosin-IIA-18; (M) mTagBFP2-PCNA-19. (N–X) N-terminal fusion constructs . (N) Cx26-mTagBFP2-7; (O) TfR-mTagBFP2-20 (transferrin receptor); (P) Golgi-mTagBFP2-7; (Q) zyxin-mTagBFP2-6;(R) VE cadherin-mTagBFP2-10; (S) mitochondria-mTagBFP2-7; (T) CENPB-mTagBFP2-22; (U) α-actinin-mTagBFP2-19; (V) c-src-mTagBFP2-7; (W) Lifeact-mTagBFP2-7; (X) vimentin-mTagBFP2-7. The cell line used for expressing mTagBFP2 fusion vectors was opossum kidney cortex proximal tubule epithelial cells (ATCC CRL-1840) in panel X, and human cervical adenocarcinoma cells (HeLa; ATCC CCL-2) in the remaining panels. The scale bar in each panel equals 10 µm.

Article Snippet: To prepare mTagBFP2 N-terminal fusions (number of linker amino acids in parenthesis), the following digests were performed: β-2 connexin-26 (7), Bam HI and Not I (rat Cx26 cDNA source: Matthias Falk, Lehigh U; NM_001004099.1); TfR (20), Bam HI and Not I (transferrin receptor cDNA source: George Patterson, NIH; NM_NM_003234); Golgi complex (7), Bam HI and Not I (human β-galactosamide α-2,6-sialyltransferase 1cDNA source: Jennifer Lippincott-Schwartz, NIH; NM_173216.2); zyxin (6), Bam HI and Not I (human zyxin cDNA source: Origene, Rockville, MD; NM_003461); vascular epithelial cadherin (10), Age I and Not I (human VE cadherin cDNA source: Origene, Rockville, MD; NM_001795.3); mitochondria (7), Bam HI and Not I (human mitochondrial targeting sequence, cytochrome c oxidase cDNA source: Clontech, Mountain View, CA; NM_004074.2); centromere protein B (22), Bam HI and Not I (human CENPB cDNA source: Alexey Khodjakov, Wadsworth Center, Albany, NY; NM_001810.5); α-actinin (19), Bam HI and Eco RI (human α-actinin cDNA source: Tom Keller, Florida State University, Tallahassee; NM_001130005.1); c-src sarcoma (7), Bam HI and Eco RI (chicken c-src cDNA source: Marilyn Resh, Sloan-Kettering, New York; XM_001232484.1); Lifeact (7), Bam HI and Not I (Lifeact cDNA source: IDT, Coralville, IA); vimentin (7), Bam HI and Not I (human vimentin cDNA source: Robert Goldman, Northwestern University; NM_003380.3).

Techniques: Construct, Expressing