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CopGFP is a recombinant green fluorescent protein (GFP) derived from the copepod Pontellina plumata. It is a naturally occurring variant of GFP and exhibits bright green fluorescence when exposed to blue or ultraviolet light.

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Lab products found in correlation

Pbabe puro mcherry egfp lc3b, by Addgene (1 mentions) Non specific shrna, by Santa Cruz Biotechnology (1 mentions) Pbabepuro gfp lc3, by Addgene (1 mentions) α tct, by Cayman Chemical (1 mentions)

3 protocols using copgfp

1

Engineered Plasmids for Autophagy Analysis

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The plasmids pBABE-puro mCherry-EGFP-LC3B and pBABEpuro GFP-LC3 (N#22418 and 22405), engineered by Jayanta Debnath [21] , were obtained from Addgene. Cell transduction and selection were performed as previously described [20] (link), [22] (link). Genetic inhibition of autophagy was achieved using ATG7 shRNA-mediated knock down (Santa Cruz). Non-specific shRNA (control) as well as copGFP were applied according to manufacturer's instructions (Santa Cruz). Transduction was performed using lentiviral particles with up to five distinct expression constructs as described elsewhere [20] (link). Transduction efficiency was quantified by the number of GFP-positive cells and in general exceeded 94% at the end of puromycin selection.
Klose J., Stankov M.V., Kleine M., Ramackers W., Panayotova-Dimitrova D., Jäger M.D., Klempnauer J., Winkler M., Bektas H., Behrens G.M, & Vondran F.W. (2014). Inhibition of Autophagic Flux by Salinomycin Results in Anti-Cancer Effect in Hepatocellular Carcinoma Cells. PLoS ONE, 9(5), e95970.
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2

Primary Rat Hippocampal Neuron Cultivation

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Primary rat hippocampal neurons were prepared from rat feti (Sprague-Dawley, day 18 of gestation; Envigo, Indianapolis, IN) as described previously [19 (link), 29 (link)–31 (link)]. Briefly, neurons (0.4 × 106 cells/ 35mm plate) were seeded on poly-l-lysin coated plates and grown in neurobasal medium supplemented with B-27, glutamine, and antibiotics (Invitrogen Gibco Life Technologies, Carlsbad, CA) for 20–22 days in vitro (DIV). Cell culture media contain 0.0916 μg/ml α-TCP with no α-TCT. α-TCT treatment: Neurons were either treated with α-TCT (1μM, ≥98% purity, Cayman chemical, Ann Arbor, MI) or a vehicle (ethanol). Neuronal media, a combination of conditioned neurobasal media with freshly prepared 1μM α-TCT or ethanol, were replaced every week for 3 weeks. Human subjects supplemented with 61.52 mg α-TCT contain α-TCT in the nanomolar range in their brain tissues [24 (link)]. Treatment with 1μM α-TCT in vitro system results in a similar range, pico to nano molar levels α-TCT in brain cells [25 (link)]. Transfection: Neurons were transfected with copGFP or lenti-Bcl-xL shRNA-GFP (Santa Cruz Biotechnology, Dallas, TX) at DIV7 and experiments were then performed 3 weeks after transfection. All protocols were approved by the Institutional Animal Care Committee (IACUC) of The University of Alabama, Tuscaloosa, AL (17-06-0324) and Yale University, New Haven, CT (2019–10388).
Park H.A., Crowe-White K.M., Ciesla L., Scott M., Bannerman S., Davis A.U., Adhikari B., Burnett G., Broman K., Ferdous K.A., Lackey K.H., Licznerski P, & Jonas E.A. (2022). Alpha-Tocotrienol Enhances Arborization of Primary Hippocampal Neurons via Upregulation of Bcl-xL. Nutrition research (New York, N.Y.), 101, 31-42.
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3

Visualizing 3D Microtissue Killing by ADP-A2M4

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To visualize killing of 3D microtissues by ADP-A2M4, A375 (MAGE-A4+ melanoma) cells were lentivirally transduced with cop-GFP (Santa Cruz Biotech). GFP+ cells were selected and maintained using puromycin and were seeded into ultra-low adhesion 384-well microplates (SBio) at 1.2 × 103 cells/well and incubated at 37°C/5% CO2 to form 3D microtissues of ~800 µm in diameter. After 6 d, transduced or non-transduced T cells were added at 2 × 104 cells/well. 3D microtissues were imaged in an Incucyte ZOOM every 4 h following target seeding. Raw fluorescent images were exported using Essen Biosciences proprietary software and microtissue area analyzed using a custom Axiovision software macro (Zeiss).
Sanderson J.P., Crowley D.J., Wiedermann G.E., Quinn L.L., Crossland K.L., Tunbridge H.M., Cornforth T.V., Barnes C.S., Ahmed T., Howe K., Saini M., Abbott R.J., Anderson V.E., Tavano B., Maroto M, & Gerry A.B. (2019). Preclinical evaluation of an affinity-enhanced MAGE-A4-specific T-cell receptor for adoptive T-cell therapy. Oncoimmunology, 9(1), 1682381.
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