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293t crl 3216

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293T (CRL-3216) is a human embryonic kidney cell line that is widely used in cell culture and molecular biology research. It is derived from 293 cells and is characterized by its high transfection efficiency and ability to produce high levels of recombinant proteins. The 293T cell line is a commonly used tool for viral vector production and gene expression studies.

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Lab products found in correlation

Human cd80 fc, by R&D Systems (2 mentions) Mouse anti human cd28 clone 28.2, by Thermo Fisher Scientific (2 mentions) Human cd86 fc, by R&D Systems (2 mentions) Alexa fluor 647 conjugated goat anti human igg secondary antibody, by Thermo Fisher Scientific (2 mentions) Jurkat e6.1 t cells, by American Type Culture Collection (2 mentions) Lncap, by American Type Culture Collection (2 mentions) Du145, by American Type Culture Collection (2 mentions) Ab6160, by Abcam (1 mentions) Anti h4, by Abcam (1 mentions) Ab1791, by Abcam (1 mentions) Anti h4k16ac, by PTM Biolabs (1 mentions) Anti tubulin, by Abcam (1 mentions) Anti sirt2, by Abcam (1 mentions) F7425, by Abcam (1 mentions) Ptm 101, by PTM Biolabs (1 mentions) Anti h3, by Abcam (1 mentions) Anti flag, by Merck Group (1 mentions) Mycoalert mycoplasma detection kit, by Lonza (1 mentions) Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Saos 2 htb 85, by American Type Culture Collection (1 mentions)

Close spelling variants of this product

CRL-3216 (164 citations) 293T CRL-3216 cells (10 citations) HEK 293T cells (ATCC CRL-3216 (6 citations) 293T (ATCC® CRL-3216™) (6 citations) HEK) 293T CRL-3216 cells (3 citations) 293T cell line (CRL-3216) (2 citations) 293t human embryonic kidney cell line (ATCC® CRL-3216™) (2 citations)

13 protocols using 293t crl 3216

1

Histone Acetylation and Deacetylation Assay

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Unless otherwise noted, all chemical reagents were purchased from Sigma-Aldrich (St. Louis, MO). D5-SB was from C/D/N Isotopes Inc., D-288. Antibodies were the following: anti-pan Kac (1:1000, PTM Biolabs, PTM-101), anti-H4K16ac (1:15,000; PTM Biolabs, PTM-122), anti-H3 (1:5000, Abcam, ab1791), anti-H4 (1:5000, Abcam, ab31830), anti-Flag (1:10,000; Sigma-Aldrich, F7425), anti-Tubulin (1:10,000; Abcam, ab6160), and anti-SIRT2 (1:1000, Abcam, 23886). Anti-pan Kbz antibody (1:1000) was made by PTM Biolabs Inc. (Chicago, IL). MEFs derived from WT and Sirt2 KO mice are kind gifts of Dr. Chu-Xia Deng (NIH)35 (link). HepG2 (HB-8065), RAW (TIB-71), and 293T (CRL-3216) cell lines were purchased from ATCC (www.atcc.org) and used without further authentication. No mycoplasma contamination was detected using a MycoAlert™ Mycoplasma Detection Kit (Lonza, LT07-118).
Huang H., Zhang D., Wang Y., Perez-Neut M., Han Z., Zheng Y.G., Hao Q, & Zhao Y. (2018). Lysine benzoylation is a histone mark regulated by SIRT2. Nature Communications, 9, 3374.
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2

Lentiviral Expression Vector Protocol

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The lentiviral expression vectors, including pLV-U6-mCherry-Puro (Cat No. VL3104), pLV-U6-EGFP-Puro (Cat No. VL3103), and pLV-EF1α-EGFP-Puro (Cat No. VL3311), were purchased from Inovogen Tech Co (Beijing, China). FLNA shRNA lentiviral particles (Cat No. sc-35374-V) and control shRNA lentiviral particles (Cat No. sc-108080) were purchased from Santa Cruz Biotech. Cell types, including SaOS2 (HTB-85, ATCC), 293T (CRL-3216, ATCC), HeLa (CCL-2, ATCC), and HepG2 (HB-8065, ATCC), were obtained from the American Type Culture Collection (ATCC) and cultured in their respective media supplied with 10% foetal bovine serum (Cat No. 10100, Gibco) and 1× penicillin–streptomycin (Cat No. 15140122, Gibco). DNA and RNA oligonucleotides were ordered from Sangon (Shanghai, China). DNA purification kits (Cat No. AP-GX-250) were purchased from Corning Co. Enzymes and transfection reagents (Cat No. 11668019) were derived from Thermo Fisher Scientific. All chemicals were obtained from Merk Co.
Wu Z., Huang L., Zhao S., Wang J., Zhang C., Song X., Chen Q., Du J., Yu D., Sun X., Zhang Y., Deng W., Zhang S, & Deng H. (2022). Early Growth Response 1 Strengthens Pol-III-Directed Transcription and Transformed Cell Proliferation by Controlling PTEN/AKT Signalling Activity. International Journal of Molecular Sciences, 23(9), 4930.
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3

Cell Line Characterization and Authentication

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IGROV-1, Vero E6 and Vero E6 TMPRSS2 clone 1 (Vero E6 TMP-1) and S-Fuse cells were described previously41 (link),35 (link). Vero E6 TMP-2 cells were kindly provided by Dr Makoto Takeda lab37 (link). 293 T (CRL-3216) and U2OS (Cat# HTB-96) cells were obtained from ATCC. Cells were authenticated by genotyping (Eurofins). Cells regularly tested negative for mycoplasma.
Planas D., Staropoli I., Michel V., Lemoine F., Donati F., Prot M., Porrot F., Guivel-Benhassine F., Jeyarajah B., Brisebarre A., Dehan O., Avon L., Bolland W.H., Hubert M., Buchrieser J., Vanhoucke T., Rosenbaum P., Veyer D., Péré H., Lina B., Trouillet-Assant S., Hocqueloux L., Prazuck T., Simon-Loriere E, & Schwartz O. (2024). Distinct evolution of SARS-CoV-2 Omicron XBB and BA.2.86/JN.1 lineages combining increased fitness and antibody evasion. Nature Communications, 15, 2254.
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4

Cell Culture of 293T and A549 Cells

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293T (CRL-3216) and A549 (CRM-CCL-185) cells were obtained from the American Type Culture Collection (ATCC) and were grown in Dulbecco’s modified Eagle medium (DMEM, Gibco) containing glutamine, supplemented with 10% fetal bovine serum (FBS). The cells were cultured at 37 °C in a humidified incubator with 5% CO2.
Tsang N.Y., Li W.F., Varhegyi E., Rong L, & Zhang H.J. (2022). Ebola Entry Inhibitors Discovered from Maesa perlarius. International Journal of Molecular Sciences, 23(5), 2620.
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5

CD28 Receptor Binding Kinetics

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293T (CRL-3216) and Jurkat T cells (E6.1) were obtained from ATCC (American Type Culture Collection). They were routinely checked for mycoplasma and were mycoplasma free. transduced with pLenti-TOPO containing wild-type CD28 and the CD28 mutants. CD28 expression was detected with a mouse anti-human CD28 (Clone 28.2; dilution 1:100; eBioscience). The binding of human CD80-Fc (catalog number 140-B1-100; R&D Systems) and human CD86-Fc (catalog number 141-B2-100; R&D Systems) to these cells were performed as previously described29 (link),58 (link). The secondary antibody used was Alexa Fluor 647 conjugated goat anti-human IgG secondary antibody (catalog number A-21445; concentration: 5 μg/ml; Life Technologies). Kd were calculated based on the presumption of one binding site. Kd= [A][B]/[AB] where in A represent soluble CD80-Fc or Cd86-Fc and B represents CD28 molecules on the cell surface of 293T cells.
Choi J., Goh G., Walradt T., Hong B.S., Bunick C.G., Chen K., Bjornson R.D., Maman Y., Wang T., Tordoff J., Carlson K., Overton J.D., Liu K.J., Lewis J.M., Devine L., Barbarotta L., Foss F.M., Subtil A., Vonderheid E.C., Edelson R.L., Schatz D.G., Boggon T.J., Girardi M, & Lifton R.P. (2015). Genomic landscape of cutaneous T cell lymphoma. Nature genetics, 47(9), 1011-1019.
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6

CD28 Receptor Binding Kinetics

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293T (CRL-3216) and Jurkat T cells (E6.1) were obtained from ATCC (American Type Culture Collection). They were routinely checked for mycoplasma and were mycoplasma free. transduced with pLenti-TOPO containing wild-type CD28 and the CD28 mutants. CD28 expression was detected with a mouse anti-human CD28 (Clone 28.2; dilution 1:100; eBioscience). The binding of human CD80-Fc (catalog number 140-B1-100; R&D Systems) and human CD86-Fc (catalog number 141-B2-100; R&D Systems) to these cells were performed as previously described29 (link),58 (link). The secondary antibody used was Alexa Fluor 647 conjugated goat anti-human IgG secondary antibody (catalog number A-21445; concentration: 5 μg/ml; Life Technologies). Kd were calculated based on the presumption of one binding site. Kd= [A][B]/[AB] where in A represent soluble CD80-Fc or Cd86-Fc and B represents CD28 molecules on the cell surface of 293T cells.
Choi J., Goh G., Walradt T., Hong B.S., Bunick C.G., Chen K., Bjornson R.D., Maman Y., Wang T., Tordoff J., Carlson K., Overton J.D., Liu K.J., Lewis J.M., Devine L., Barbarotta L., Foss F.M., Subtil A., Vonderheid E.C., Edelson R.L., Schatz D.G., Boggon T.J., Girardi M, & Lifton R.P. (2015). Genomic landscape of cutaneous T cell lymphoma. Nature genetics, 47(9), 1011-1019.
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7

Cell Line Culturing and Reagent Procurement

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Hep3B (HB-8064), Hepa1-6 (CRL-1830), and 293 T (CRL-3216) cell lines were purchased from the American Type Culture Collection (ATCC). Huh7 cell line (JCRB0403) was purchased from the Japanese Cancer Research Resources Bank (JCRB). H22 cell line (GDC0091) was purchased from China Center for Type Culture Collection (CCTCC). Cells were routinely cultured in Dulbecco’s modified Eagle medium (DMEM) containing 10% fetal bovine serum (FBS) (Wisent, Nanjing, China) at 37 °C. MG132 (S2619) and cycloheximide (CHX) were purchased from Sigma-Aldrich.
Li Q., Zhang L., You W., Xu J., Dai J., Hua D., Zhang R., Yao F., Zhou S., Huang W., Dai Y., Zhang Y., Baheti T., Qian X., Pu L., Xu J., Xia Y., Zhang C., Tang J, & Wang X. (2022). PRDM1/BLIMP1 induces cancer immune evasion by modulating the USP22-SPI1-PD-L1 axis in hepatocellular carcinoma cells. Nature Communications, 13, 7677.
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8

Culturing Human Prostate Cell Lines

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Human prostate cancer cell lines (VCaP, PC3, 22RV1, DU145 and LNCaP), human prostate epithelial cell line (RWPE) and 293 T(CRL-3216) were obtained from the American Type Culture Collection (Rockville, MD, USA) and cultured following the manufacturer’s recommendations. Detailed information of protocols for transient and stable transfection are available in Additional file 2: Materials and Methods.
Hu J., Sun F., Chen W., Zhang J., Zhang T., Qi M., Feng T., Liu H., Li X., Xing Y., Xiong X., Shi B., Zhou G, & Han B. (2019). BTF3 sustains cancer stem-like phenotype of prostate cancer via stabilization of BMI1. Journal of Experimental & Clinical Cancer Research : CR, 38, 227.
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9

Prostate Cancer Cell Line Authentication and Transfection

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Human PCa cell lines (VCaP, PC3, 22RV1, DU145 and LNCaP), human prostate epithelial cell line (RWPE) and 293T (CRL-3216) were obtained from the American Type Culture Collection (Rockville, MD, USA) between 2012 and 2015 and authenticated again by short tandem repeat analysis again before and after our study. The cumulative culture length of the cells between thawing and use in this study was less than 15 passages. All of the newly revived cells were tested free of mycoplasma contamination by Hoechst 33258 staining (Beyotime, Jiangsu, China). Cells were transfected with miR-204 mimic/inhibitor, SOX4 siRNA (GenePharma, Shanghai, China), CUL4B siRNA 1#, 2# (Qiagen, Hilden, Germany), EZH2 siRNA (GenePharma, Shanghai, China) or the respective negative controls, using Hiperfect transfection reagent (Qiagen). CUL4B Plasmids were transfected with Lipofectamine 2000 (Invitrogen, Carlsbad, USA) according to the manufacturer’s instructions. For stable knockdown or overexpression of CUL4B, Lenti-shCUL4B-GFP, Lenti-CUL4B-Flag as well as their controls (Lenti-vector control and Lenti-shSCR) were transfected into VCaP or DU145 cells. The overexpression sequences and targeted sequences for siRNAs and shRNAs were described in Supplementary Table 2.
Qi M., Hu J., Cui Y., Jiao M., Feng T., Li X., Pang Y., Chen X., Qin R., Su P., Zhang H., Wang Y., Gong Y, & Han B. (2019). CUL4B promotes prostate cancer progression by forming positive feedback loop with SOX4. Oncogenesis, 8(3), 23.
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10

Obtaining 293 T Cell Line

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293 T (CRL-3216™) were obtained from American Type Culture Collection (ATCC), suggested to be female in origin.
Dahiya S., Beier U.H., Wang L., Han R., Jiao J., Akimova T., Angelin A., Wallace D.C, & Hancock W.W. (2020). HDAC10 deletion promotes Foxp3+ T-regulatory cell function. Scientific Reports, 10, 424.
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