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Slc1a5

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SLC1A5 is a sodium-dependent neutral amino acid transporter that mediates the transport of neutral amino acids, such as glutamine, across the cell membrane. It plays a crucial role in cellular metabolism and various biological processes.

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Lab products found in correlation

Glutaminase, by Abcam (2 mentions) Smad2 3, by Santa Cruz Biotechnology (2 mentions) Phospho akt, by Santa Cruz Biotechnology (2 mentions) Cxcr4, by Abcam (1 mentions) Glyceraldehyde 3 phosphate dehydrogenase gapdh, by R&D Systems (1 mentions) Phospho smad2 3, by Santa Cruz Biotechnology (1 mentions) Phospho src, by Santa Cruz Biotechnology (1 mentions) Sdf 1α, by Abcam (1 mentions) Cd62p, by Abcam (1 mentions) β catenin, by Santa Cruz Biotechnology (1 mentions) T per, by Thermo Fisher Scientific (1 mentions) Image master id, by GE Healthcare (1 mentions) Cyclin d1, by Santa Cruz Biotechnology (1 mentions) Protease inhibitor, by Beyotime (1 mentions) β actin, by CWBIO (1 mentions) Phgdh, by Proteintech (1 mentions) C myc, by Proteintech (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Slc7a5, by Santa Cruz Biotechnology (1 mentions)

6 protocols using slc1a5

1

Comprehensive Protein Expression Analysis

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Cells and resected tumor tissues were subjected to protein extraction using tissue protein reagents (T-PER, Pierce Biotechnology, Rockford, IL, USA). Proteins were separated by SDS-PAGE and then transferred to PVDF membranes. The membranes were incubated with the following antibodies: cyclin D1, β-catenin, Src, phospho-Src, Akt, phospho-Akt, Smad2/3, phospho-Smad2/3, Thromboxane A2 Receptor (TXA2R) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), Glucose Transporter-1 (GLUT1), PKM2, SLC1A5, SLC7A5, glutaminase, CD41, CD62P, CD45, SDF-1α, CXCR4 (Abcam, Cambridge, MA, USA), and Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) (R&D Systems, Minneapolis, MN, USA). The reacted proteins were further determined with horseradish peroxidase-labeled IgG, visualized using Enhanced Chemiluminescence (ECL) Western blotting reagents, and quantified by the optical densitometry (Image Master ID, Pharmacia Biotech, Upsalla, Sweden) of developed autoradiographs.
Wang J.D., Chen W.Y., Li J.R., Lin S.Y., Wang Y.Y., Wu C.C., Liao S.L., Ko C.C, & Chen C.J. (2020). Aspirin Mitigated Tumor Growth in Obese Mice Involving Metabolic Inhibition. Cells, 9(3), 569.
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2

Protein Expression Analysis by Western Blot

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Cells were harvested in RIPA lysis buffer (Beyotime, Shanghai, China) containing protease inhibitors (Beyotime, Shanghai, China) for 10 mins, and the lysates were centrifuged at 12,000 rpm for 10 mins at 4°C. Then, the supernatants were collected to determine the protein concentrations using a BCA Protein Assay Kit (Beyotime, Shanghai, China). Afterwards, these proteins were separated using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto a polyvinylidene fluoride (PVDF) membrane that was activated in advance by methanol. After blocking with 5% non-fat dry milk in Tris-buffered saline with Tween (TBST) at room temperature for one hour, these membranes were incubated with the primary antibodies overnight against SLC1A5 (Abcam, MA, USA), GLS (Proteintech, Wuhan, China), c-Myc (Proteintech, Wuhan, China), β-actin (CWBiotech, Beijing, China), PHGDH (Proteintech, Wuhan, China), and PSPH (Proteintech, Wuhan, China). Then, these membranes were incubated with secondary antibodies (CWBiotech, Beijing, China) the following day. After incubation, the membrane was scanned.
Zhang P., Wang Q., Lin Z., Yang P., Dou K, & Zhang R. (2019). Berberine Inhibits Growth of Liver Cancer Cells by Suppressing Glutamine Uptake. OncoTargets and therapy, 12, 11751-11763.
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3

Quantitative Protein Analysis Workflow

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Whole steps of tissue protein extraction, cell lysate preparation, SDS-PAGE, Western blotting and quantitative calculation were performed as reported procedures [32 (link),33 (link)]. Interesting molecules corresponding to the indicated antibodies were cyclin D1 (sc-56302), β-catenin (sc-133240), HMGB1 (sc-135809), RAGE (sc-74535), Akt (sc-5298), phospho-Akt (sc-293125), SLC7A5 (sc-374232), Smad2/3 (sc-398844), phospho-Smad2/3 (sc-11769) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), GLUT1 (ab115730), PKM2 (ab137852), SLC1A5 (ab84903), glutaminase (ab150474), CD62P (ab59738), MPO (ab65871) (Abcam, Cambridge, MA, USA) and Glyceraldehyde-3-Phosphate Dehydrogenase (AF5718, GAPDH) (R&D Systems, Minneapolis, MN, USA).The proteins of interest were visualized and quantified using a G:BOX mini multi-fluorescence and chemiluminescence imaging system (Syngene, Frederick, MD, USA).
Chen C.J., Wu C.C., Chang C.Y., Li J.R., Ou Y.C., Chen W.Y., Liao S.L, & Wang J.D. (2022). Metformin Mitigated Obesity-Driven Cancer Aggressiveness in Tumor-Bearing Mice. International Journal of Molecular Sciences, 23(16), 9134.
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4

Apoptosis pathway inhibitors protocol

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ABT-263, A-1331852 and A-1210477 were from AbbVie (North Chicago, IL, USA),
ABT-199, epigallocatechin gallate (EGCG), CB-839, simvastatin, rapamycin and
torin-1 from Selleck Chemicals (Houston, TX, USA),
gamma-L-glutamyl-p-nitroanilide (GPNA) from Insight
Biotechnology (Wembley, Middlesex, UK), azaserine from Cambridge Bioscience
(Cambridge, UK), aminooxyacetate (AOA), sodium palmitate, dimethyl
α-ketoglutarate, oxaloacetate and citrate from Sigma-Aldrich
(Gillingham, UK), L-glutamine from Life Technologies (Paisley, UK) and
GSK2194069, SB204990, atorvastatin, pitavastatin and bafilomycin A1 from Tocris
(Abingdon, UK). Antibodies against PARP, BCL-2, MCL-1, BAX, BAK and GAPDH were
from Santa Cruz Biotechnology (Santa Cruz, CA, USA), caspase-3, caspase-9,
BCL-XL, BCL-w, BIM, PUMA, BAD, IDH2, ACL, ACO2, ATG5 and ATG7
from Cell Signaling Technology (MA, USA), BID from Prof. J. Borst (The
Netherlands Cancer Institute, Amsterdam, the Netherlands), NOXA from Millipore
(Watford, UK) and SLC1A5, GLS, GFAT, GLUD1, IDH3, FASN and HMGR from Abcam
(Cambridge, UK).
Al-Zebeeby A., Vogler M., Milani M., Richards C., Alotibi A., Greaves G., Dyer M.J., Cohen G.M, & Varadarajan S. (2019). Targeting intermediary metabolism enhances the efficacy of BH3 mimetic therapy in hematologic malignancies. Haematologica, 104(5), 1016-1025.
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5

Visualizing GSC-COL1 Interactions in GBM

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For immunostaining analysis of the contact of GSCs to COL1, GBM sections were fixed with 4% PFA for 20 minutes at room temperature, washed three times with PBS, and then blocked with a PBS‐based solution containing 5% normal goat serum and 0.3% Triton X‐100. Cells were co‐incubated overnight at 4 °C with goat polyclonal anti‐COL1A1 antibody (Abcam) or SLC1A5 (CST) and mouse monoclonal anti‐CD133 (W6B3C1 clone) (Miltenyi Biotec). After washed three times with PBS, cells were co‐incubated with Alexa 488 conjugated donkey anti‐mouse IgG (Invitrogen; 1:400) and Alexa 594 conjugated donkey anti‐rabbit IgG (Invitrogen, 1:800). Nucleus were counterstained with DAPI (Sigma; 10 µg ml−1). Immunofluorescent images were collected on a Leica TCS SP5 confocal microscope and analyzed using LAS AF software.
Wei Y., Geng S., Si Y., Yang Y., Chen Q., Huang S., Chen X., Xu W., Liu Y, & Jiang J. (2023). The Interaction between Collagen 1 and High Mannose Type CD133 Up‐Regulates Glutamine Transporter SLC1A5 to Promote the Tumorigenesis of Glioblastoma Stem Cells. Advanced Science, 11(3), 2306715.
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6

Correlation of circ_0000808 Expression and NSCLC

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A total of 63 patients with NSCLC were recruited from the Shanghai Pulmonary Hospital, Tongji University School of Medicine, and their NSCLC tumor tissues and adjacent normal tissues were collected and stored at −80 °C. The clinicopathologic features in NSCLC patients were shown in Table 1. A part of fresh tissue was taken to prepare paraffin sections to perform immunohistochemical (IHC) staining using SP Kit (Invitrogen, Carlsbad, CA, USA) and anti-solute carrier family 1 member 5 (SLC1A5) (1:1600, Abcam, Cambridge, MA, USA). For this study, all patients signed written informed consent and received the approval from the Ethics Committee of the Shanghai Pulmonary Hospital, Tongji University School of Medicine.

Correlation of the expression of circ_0000808 with clinicopathologic features in NSCLC patients

ParametersN = 63circ_0000808 expressionp-value
High N = 31Low N = 32
Age, years
 ≤ 602512130.719
 > 60382018
Sex
 Male4321220.649
 Female20119
Smoking history
 Yes3920190.921
 No241212
Tumor location
 Left lobe2715120.513
 Right lobe361719
Tumor size
 ≤ 33010200.008
 > 3332211
TNM stage
 I + II stage4921280.018
 III stage14113
Lymph node metastasis
 No431528< 0.001
 Yes20173
Cai Y., Dong Z, & Wang J. (2022). Circ_0000808 promotes the development of non-small cell lung cancer by regulating glutamine metabolism via the miR-1827/SLC1A5 axis. World Journal of Surgical Oncology, 20, 329.
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