st6gal2  (R&D Systems)

Journal: Cell Proliferation

Article Title: Glycomics reveal that ST6GAL1‐mediated sialylation regulates uterine lumen closure during implantation

doi: 10.1111/cpr.13169

Figure Lengend Snippet: Expression patterns of the sialylated glycans as well as the mRNA and protein expression of ST6GAL1 and ST6GAL2 in pig endometrium during implantation. (A) The 19 possible α2,3/6‐sialylated N‐glycans identified by MALDI‐TOF MS. Fucose ; N‐acetylglucosamine ; N‐acetyl neuraminic acid ; N‐glycolyl neuraminic acid ; mannose ; Galactose . (B) Representative images of lectin fluorescence assays taken from pig endometrium on gestational days 12, 15 and 18 (n = 3 gilts/gestational day). MAL‐II was used to detect α2,3‐linked sialic acid residues. SNA was used to detect α2,6‐linked sialic acid residues. (C) Expression levels of ST6GAL1 and ST6GAL2 in pig endometrium on gestational days 12, 15 and 18 measured by qRT‐PCR (n = 3 gilts/gestational day). The error bars represent the standard error. Mean is denoted by a red line. Nonparametric Mann‐Whitney one tail test was used for statistical analysis. (D) Distributions of ST6GAL1 and ST6GAL2 in pig endometrium on gestational days 12, 15 and 18 (n = 3 gilts/gestational day). The positive signal is in green, while the nucleus is in blue. NC, negative control. LE, endometrial luminal epithelium. GD, gestational day. Scale bar = 50 µm

Article Snippet: The primary antibodies used were ST6GAL1 (1:50, ab77676, abcam), ST6GAL2 (1:50, AF7747; R&D Systems), E‐cadherin (1:50, ab40772; abcam), β‐catenin (1:50, ab6302; abcam), Vimentin (1:50; sc‐73258), Rac1 (1:30, 66122–1‐Ig; Proteintech) and RhoA (1:50, 10749–1‐Ig; Proteintech).

Techniques: Expressing, Fluorescence, Quantitative RT-PCR, MANN-WHITNEY, Negative Control

Journal: Journal of Cancer

Article Title: ASPN and GJB2 Are Implicated in the Mechanisms of Invasion of Ductal Breast Carcinomas

doi: 10.7150/jca.4120

Figure Lengend Snippet: Primary Antibodies and other conditions for immunohistochemical analysis

Article Snippet: ST6GAL2 , Polyclonal , 1:20 , Sigma , PT link pH 6..

Techniques: Immunohistochemical staining

Journal: Journal of Cancer

Article Title: ASPN and GJB2 Are Implicated in the Mechanisms of Invasion of Ductal Breast Carcinomas

doi: 10.7150/jca.4120

Figure Lengend Snippet: Validation of microarray-based gene expression results by qRT-PCR.

Article Snippet: ST6GAL2 , Polyclonal , 1:20 , Sigma , PT link pH 6..

Techniques: Microarray, Expressing

Journal: Journal of Cancer

Article Title: ASPN and GJB2 Are Implicated in the Mechanisms of Invasion of Ductal Breast Carcinomas

doi: 10.7150/jca.4120

Figure Lengend Snippet: Expression of ASPN, GJB2, ENPP2, ST6GAL2, and TMSB10 genes. (A) Histograms showing differential gene expression between DCIS pure vs normal tissue, DCIS associate to IDC vs normal breast tissue, IDC vs normal tissue, DCIS associate to IDC vs DCIS pure samples and IDC vs DCIS pure samples. Gene expression levels of ASPN, GJB2, ENPP2, ST6GAL2 and TMSB10 were determined by qRT-PCR. The bar represents the mean ± S.E. of all samples by duplicate. Asterisks (*) indicates statistically significant differences ( P < 0.05). (B) Immunohistochemical images of asporin and GJB2 (connexin-26) expressions in normal tissue (black arrow), proliferative non-invasive (two black arrows) and invasive neoplasms (black star).

Article Snippet: ST6GAL2 , Polyclonal , 1:20 , Sigma , PT link pH 6..

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining

Journal: Journal of Cancer

Article Title: ASPN and GJB2 Are Implicated in the Mechanisms of Invasion of Ductal Breast Carcinomas

doi: 10.7150/jca.4120

Figure Lengend Snippet: Microscopic images and qRT-PCR of mice tumors. (A) Microscopic images of typical breast tissue in MMTV-PyMT mice at ages of 0 weeks (considered as normal tissue), 8 weeks (equivalents to DCIS), and in 13 weeks (equivalent to IDC). Tumor proliferation overgrowth stroma with the time as the grade of nuclear atypia increases. Mitoses are clearly found in IDC (see arrow), H&E staining, x40 magnification. (B) Histograms showing relative fold changes in gene expression by qRT-PCR of ASPN, GJB2, and ST6GAL2. Significative increased overexpression of GJB2 in DCIS and IDC with respect to normal tissue. Each bar represents the mean ± S.E. of all samples by duplicate. Asterisk (*) denote statistically significant differences ( P < 0.05).

Article Snippet: ST6GAL2 , Polyclonal , 1:20 , Sigma , PT link pH 6..

Techniques: Quantitative RT-PCR, Staining, Expressing, Over Expression

Journal: Nature Communications

Article Title: Loss of α2-6 sialylation promotes the transformation of synovial fibroblasts into a pro-inflammatory phenotype in arthritis

doi: 10.1038/s41467-021-22365-z

Figure Lengend Snippet: a Illustration shows pathways involved in sialic acid and sialylated glycans biosynthesis. Relative expression for each gene was extracted from bulk RNA-Seq data comparing naive and arthritic SFs directly isolated from mouse synovium. b Synovial fibroblasts expanded from healthy or arthritic mouse joints were stimulated in vitro for 6 h with recombinant IL-1β, TNF, or IL-17A [10 ng/ml]. mRNA expression for genes involved in sialylation [as shown in a ] was evaluated by RT-qPCR. Il6 was included as a positive control to confirm cell activation. Volcano plots show log2 fold difference in stimulated cells ( x axis) and p -value ( y axis). Each dot represents the mean of three independent experiments analyzed in triplicate, with naive cells in blue and arthritic cells in red. The pink line indicates a twofold-change in gene expression threshold and blue lines indicate p = 0.05 threshold in the t -test to evaluate statistical significance. c TNF, but not IL-1β or IL-17A reduces α2-6-sialylation of synovial fibroblasts both at RNA ( St6gal1 expression) and protein (SNA binding) levels. Expression of St6gal1 (left panel ) was evaluated by RT-qPCR. Results show the mean of three independent experiments analyzed in triplicate, error bars represent SEM. Statistical significance was determined using two-tail unpaired t -tests where * p < 0.05 and ** p < 0.01. Actual p -values: a 0.0069, b 0.0277. Expression of α2-6 sialylated glycoconjugates was determined by SNA binding (right panel) as in Fig. . Results are merged from 3 (naive) and 2 (CIA) independent experiments, n = 10 (DMEM naive), 16 (IL-1 naive and IL-17 naive), 13 (TNF naive), 10 (DMEM CIA), 12 (IL-1 naive and IL-17 naive), and 9 (TNF CIA). Statistical significance was determined using two-tail unpaired t -tests, where ** p < 0.01 and *** p < 0.001. Actual p -values: c 0.0124, d 0.0091. d SFs were expanded from OA human synovium. Cells were stimulated with recombinant human TNF (10 ng/ml) for 6 h. IL6 , ST6GAL1, and ST6GAL2 expression were determined by qPCR. Results show relative expression to HPRT, showing mean ± SEM. Statistics: one-tail unpaired t -test, n = 3 biological replicates from cells pooled from three donors, *** p < 0.001, * p < 0.05. Actual p -values: e <0.0001, f 0.0170, g 0.0321. e Relative expression of individual sialylated glycan structures was evaluated by MALDI-TOF MS analysis as in Fig. . Ratios of sialylated vs non-sialylated twin structures were evaluated for non-stimulated cells (blue) and TNF stimulated (red) cells (48 h, 10 ng/ml). Results are from one single experiment using pooled cells from three animals.

Article Snippet: TaqMan predesigned probes (ThermoFisher Scientific) were used to evaluate mouse Actb (4352933E), Il6 (Mm00446190_m1 ), St6gal1 (Mn00486119_m1) , St6gal2( Mm01268915_m1) , St3gal1 (Mm00501493_m1) , St6galnac1 (Mm01252949_m1) , St6galnac3 (Mm01316813_m1) and Mmp3 (Mm00440295_m1) and human IL6( Hs00174131_m1) , ST6GAL1 (Hs00949382_m1) , ST6GAL2 (Hs00383641) and HPRT (4333768T).

Techniques: Expressing, RNA Sequencing, Isolation, In Vitro, Recombinant, Quantitative RT-PCR, Positive Control, Activation Assay, Gene Expression, Binding Assay, Glycoproteomics