Journal: Molecular & Cellular Proteomics : MCP

Article Title: Comparative Tissue Proteomics of Microdissected Specimens Reveals Novel Candidate Biomarkers of Bladder Cancer *

doi: 10.1074/mcp.M115.051524

Figure Lengend Snippet: Correlations of SLC3A2, STMN1, and TAGLN2 protein expression (IHC scores) with clinicopathological characteristics in tissue specimens

Article Snippet: The urinary concentrations of the four candidate proteins were measured using commercial ELISA kits for SLC3A2 (Cusabio, Wuhan, China), SFN (Cusabio, Wuhan, China) and STMN1 (USCN Life Sciences, Wuhan, China), and an ELISA developed in house for TAGLN2.

Techniques: Expressing

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Comparative Tissue Proteomics of Microdissected Specimens Reveals Novel Candidate Biomarkers of Bladder Cancer *

doi: 10.1074/mcp.M115.051524

Figure Lengend Snippet: Details of biomarker discovery data and verification of seven selected protein candidates in tissue and urine specimens

Article Snippet: The urinary concentrations of the four candidate proteins were measured using commercial ELISA kits for SLC3A2 (Cusabio, Wuhan, China), SFN (Cusabio, Wuhan, China) and STMN1 (USCN Life Sciences, Wuhan, China), and an ELISA developed in house for TAGLN2.

Techniques: Biomarker Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Marker

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Comparative Tissue Proteomics of Microdissected Specimens Reveals Novel Candidate Biomarkers of Bladder Cancer *

doi: 10.1074/mcp.M115.051524

Figure Lengend Snippet: Up-regulation of A, SLC3A2; B, STMN1; and C, TAGLN2 in individual bladder tumor tissue specimens determined by IHC.

Article Snippet: The urinary concentrations of the four candidate proteins were measured using commercial ELISA kits for SLC3A2 (Cusabio, Wuhan, China), SFN (Cusabio, Wuhan, China) and STMN1 (USCN Life Sciences, Wuhan, China), and an ELISA developed in house for TAGLN2.

Techniques:

Journal: Frontiers in Oncology

Article Title: Decrease of Intracellular Glutamine by STF-62247 Results in the Accumulation of Lipid Droplets in von Hippel-Lindau Deficient Cells

doi: 10.3389/fonc.2022.841054

Figure Lengend Snippet: Effects of STF-62247 on glutamine transporters. (A, B) mRNA relative expression, measured by RT-qPCR, of glutamine transporters SLC1A5, SLC7A5 and SLC3A2 influenced by (A) VHL status and (B) STF-62247. RCC4 VHL- and VHL+ were treated with 1.25 µM STF-62447 for 24 hr. Results for treated cells are compared to control cells (represented by the dotted line at 1). Results are presented as means and SEM of three independent experiments. (C) Glutamine transporters, SLC1A5, SL7A5 and SLC3A2, protein levels in RCC4 VHL- and VHL+ treated to STF-62247 for 24 and 48 hr. (D) Intracellular leucine was measured by LC-MS in RCC4 VHL- and VHL+ treated with 1.25 µM STF-62247 for 24 hr (N=6). Student’s t -tests were performed to compare results between VHL- and VHL+ cells (A) or between controls and treated cells (B, D) (*p < 0.05, **p < 0.01, ***p < 0.001). (E) Cell viability was evaluated by XTT assay in RCC4 VHL- and VHL+ cells. SLC1A5 and SLC7A5 inhibitors, GPNA and BCH respectively, were tested alone (concentrations from 0 to 1 mM) or combined with 1.25 µM STF-62247 (N=3). Two-Way ANOVA followed by Tukey’s test was performed to assess statistically significant results. Comparison between CTL and STF-62247 conditions for each concentration of inhibitor (GPNA or BCH) are denoted with the following statistical marks *p < 0.05, **p < 0.01 or ***p < 0.001. Comparison of each inhibitor concentrations (x-axis) are made with their respective control (0 mM columns, with or without STF-62247) and statistical significances are denoted by # p < 0.05, ## p < 0.01 or ### p < 0.001.

Article Snippet: Membranes were blocked using 5% skim milk diluted in a solution of 0.075% PBS-Tween (PBS-T) and incubated overnight in 3% BSA with specific primary antibodies against VHL, HIF-1α and HIF-2α (Cell signaling #68547, 14179, 59973), SLC1A5 and β-actin (Santa Cruz Biotechnologies sc-99002 and sc-47778), SLC7A5 (Medical & Biological Laboratories #BMP011), SLC3A2 (Aviva Systems Biology #OAAB00158), ASNS (Signalway #32909), SCD1 (Applied Biological Materials #ABM-G076), PLIN2 (Proteintech #15294-1-AP) and CPT1A (Abcam #ab220789).

Techniques: Expressing, Quantitative RT-PCR, Control, Liquid Chromatography with Mass Spectroscopy, XTT Assay, Comparison, Concentration Assay

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Endothelial Cells Require CD98 for Efficient Angiogenesis—Brief Report

doi: 10.1161/atvbaha.116.308335

Figure Lengend Snippet: Figure 1. Endothelial CD98hc is critical for angiogenesis. Subcutaneous B16 tumors were grown in VE-Cad-CreERT2Slc3a2fl/fl and Slc3a2fl/fl mice treated with tamoxifen for 5 d before for (A–C). A, CD98hc expression on tumor endothelial cells (ECs). Tumors were harvested and collagenase digested from B16 melanoma-bearing mice on day 21 after tumor cell inoculation. ECs were enriched with CD31-coated beads and stained with antibodies for mouse CD98hc (or isotype control) on CD31+ CD102+ cells, followed by flow cytom- etry. B, Tumor growth after EC-specific CD98 deletion. B16 tumors were measured every 2 d beginning on day 7 (left). On day 18, mice were euthanized, and excised tumors weighed (right). Error bars=SEM for n=10 mice per group. *P<0.05 (1-tailed t test). C, Tumor vasculature. B16 tumor sections were stained for CD31; representative images are shown on left with summary quantification from >25 image fields on right. Scale bar=250 μm. D, Retinal angiogenesis. VE-Cad-CreERT2Slc3a2fl/fl and Slc3a2fl/fl mice littermate mice were treated daily with tamoxifen on postnatal day 3 (P3) to P5 and euthanized on P10. Retinas were stained for CD98hc and blood vessels (isolectin B4); representative 2-channel close-up images of whole mount retinas with a ×20 objective are shown. Multiple retinal regions were computer-merged to create whole-mount images (E). Bar in (D)=50 μm; bar in (E)=500 μm. F, Retinal vessel coverage. Summary bar graph shows areas covered by blood vessel staining in merged images from (E) that were calculated using ImageJ software. Error bars are SEM for n=4 per group.

Article Snippet: Rag1-/-, and Rosa-26(Cre-ER) mice were obtained from Jackson Labs, VE-Cad-CreER T2 mice 1 are from Taconic, and the CD98 heavy chain conditional (Slc3a2 fl/fl ) strain was generated as previously described 2 .

Techniques: Expressing, Staining, Control, Software