Journal: iScience

Article Title: Perturbation of placental protein glycosylation by endoplasmic reticulum stress promotes maladaptation of maternal hepatic glucose metabolism

doi: 10.1016/j.isci.2022.105911

Figure Lengend Snippet:

Article Snippet: Datura stramonium Agglutinin-biotinylated lectin , Vector Laboratories , Cat#B-1185.

Techniques: Recombinant, Plasmid Preparation, Avidin-Biotin Assay, Lysis, Isolation, Bradford Protein Assay, Labeling, SYBR Green Assay, Mutagenesis, Software

Journal: iScience

Article Title: Perturbation of placental protein glycosylation by endoplasmic reticulum stress promotes maladaptation of maternal hepatic glucose metabolism

doi: 10.1016/j.isci.2022.105911

Figure Lengend Snippet:

Article Snippet: After washing in water, sections were incubated in 10 μm/mL biotinylated lectin (Concanavalin A, P. sativum and D. stramonium Agglutinins (B-1005, B-1055, B-1185) Vector Laboratories, Peterborough, UK) in 50 mM TBS containing 1 mM calcium chloride for 75 min at 37°C, washed 3 times 5 min in the same buffer, then treated with 5 μg/mL avidin peroxidase (A3151, Sigma-Aldrich) in 125 mM TBS, pH 7.6, with added 347 mM sodium chloride for 90 min at 37°C.

Techniques: Recombinant, Plasmid Preparation, Avidin-Biotin Assay, Lysis, Isolation, Bradford Protein Assay, Labeling, SYBR Green Assay, Mutagenesis, Software

Journal: PLoS ONE

Article Title: β1,6 GlcNAc Branches-Modified PTPRT Attenuates Its Activity and Promotes Cell Migration by STAT3 Pathway

doi: 10.1371/journal.pone.0098052

Figure Lengend Snippet: (A) The level of GnT-V protein was detected in different cell lines by immunoblot. (B) The GnT-V-7721 or GnT-V-HT29 stable cells were identified by detecting the level of GnT-V protein using immunoblot. (C) GnT-V-7721 and Mock-7721 cells were seeded on coverslips, followed by fixation and staining with biotinylated L-PHA and FITC-conjugated avidin, and then visualized under fluorescence microscopy. (D) Flow cytometry was performed with biotinylated L-PHA in GnT-V-7721 and GnT-V-HT29 stable cells, as well as Mock cells. FITC-conjugated avidin D staining was used as negative control.

Article Snippet: Biotinylated phaseolus vulgaris leucoagglutinin (L-PHA), biotinylated datura stramonium lectin (DSL), agarose bound L-PHA, agarose bound streptavidin and fluorescein anti-avidin D were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Western Blot, Staining, Avidin-Biotin Assay, Fluorescence, Microscopy, Flow Cytometry, Negative Control

Journal: PLoS ONE

Article Title: β1,6 GlcNAc Branches-Modified PTPRT Attenuates Its Activity and Promotes Cell Migration by STAT3 Pathway

doi: 10.1371/journal.pone.0098052

Figure Lengend Snippet: (A) 4 types of RPTPs were evaluated in Mock-7721 and GnT-V-7721 using immunofluorescent staining. Cells were placed on coverslips overnight and stained with anti-PTPRM, anti-PTPRT, anti-PTPRK, or anti-PTPRU antibody, respectively. Then, FITC-labeled anti-mouse IgG or Cy3-labeled anti-rabbit IgG was used based on the specificity of the primary antibody. Intensity of fluorescent was visualized under confocal microscopy. (B) Lectin precipitation was performed with L-PHA bounded agarose, followed by immunoblot with anti-PTPRT antibody in GnT-V-7721 and Mock-7721. (C) PTPRT was immunoprecipitated in GnT-V-7721 and Mock-7721, and then subjected to the immunoblot with biotinylated L-PHA, biotinylated DSL and anti-PTPRT antibody, respectively.

Article Snippet: Biotinylated phaseolus vulgaris leucoagglutinin (L-PHA), biotinylated datura stramonium lectin (DSL), agarose bound L-PHA, agarose bound streptavidin and fluorescein anti-avidin D were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Staining, Labeling, Confocal Microscopy, Western Blot, Immunoprecipitation

Journal: PLoS ONE

Article Title: β1,6 GlcNAc Branches-Modified PTPRT Attenuates Its Activity and Promotes Cell Migration by STAT3 Pathway

doi: 10.1371/journal.pone.0098052

Figure Lengend Snippet: (A) Confocal microscopy was performed to observe the distribution of PTPRT in Mock-7721 and GnT-V-7721 cells. The mouse anti-human PTPRT and FITC-conjugated goat anti-mouse IgG were used. The bottom panel represents the magnified images of indicated area in the top panel. (B) Cell-surface retention of PTPRT was examined after cell-surface protein was biotinylated. The stable transfectants were re-cultured for 3 and 6 hours before they were homogenized and pulled down by streptavidin bounded agarose precipitation. After the precipitation, PTPRT was detected by immunoblot using anti-PTPRT antibody. The bottom chart is the quantification of band intensity in the upper panel. The band intensity of PTPRT at 3 or 6 hour time point was normalized to that at 0 hour in both Mock-7721 and GnT-V-7721. The data represent the mean ± SEM of three independent analyses. (C) Mock-7721, GnT-V-7721, Mock-HT29 and GnT-V-HT29 cells were grown to confluence in 6-well dishes and cross-linked with 3 mM BS 3 at 4°C for 2 hours, followed by immunoblot for PTPRT. The graph (right panel) is the band intensity analysis of dimer band intensity normalized to its total monomer. The data represent the mean ± SEM of three independent analyses.

Article Snippet: Biotinylated phaseolus vulgaris leucoagglutinin (L-PHA), biotinylated datura stramonium lectin (DSL), agarose bound L-PHA, agarose bound streptavidin and fluorescein anti-avidin D were purchased from Vector Laboratories (Burlingame, CA).

Techniques: Confocal Microscopy, Cell Culture, Western Blot