Plasmid Transfection in HeLa Cells

The following plasmids have been described previously: pC97ELsLuc [36 (link), 37 ], pBELsLuc [36 (link), 37 ], pHPV16AN [36 (link), 37 ], and pX856F [38 (link)]. PcDNA3-FLAG-HA-hYTHDC1 (#85167), pcDNA3/Flag-METTL3 (#53739), pcDNA3/Flag-METTL14 (#53740), pcDNA3/Flag-WTAP (#53741), pFRT/TO/HIS/FLAG/HA-ALKBH5 (#38073) were purchased from Addgene. pcDNA3.1⁺/C-(K)DYK-FTO was purchased from GenScript. Transfections were made with Turbofect according to the manufacturer’s protocol (Fermentas). Briefly, a mixture of 2 µl Turbofect per 1 µg DNA and 100 µl of DMEM without serum was incubated at room temperature for 25 min prior to dropwise addition to 60-mm plates with subconfluent HeLa cells.

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Cui X., Nilsson K., Kajitani N, & Schwartz S. (2022). Overexpression of m6A-factors METTL3, ALKBH5, and YTHDC1 alters HPV16 mRNA splicing. Virus Genes, 58(2), 98-112.

Publication 2022

pcDNA3/Flag-METTL3 pcDNA3/Flag-METTL14 Turbofect

A dna Hela cells Mettl14 Mettl3 Per 1 Plasmids Serum Transfections

Corresponding Organization :

Other organizations : Lund University, Uppsala University

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Variable analysis

independent variables

PC97ELsLuc
PBELsLuc
PHPV16AN
PX856F
PcDNA3-FLAG-HA-hYTHDC1
PcDNA3/Flag-METTL3
PcDNA3/Flag-METTL14
PcDNA3/Flag-WTAP
PFRT/TO/HIS/FLAG/HA-ALKBH5
PcDNA3.1+/C-(K)DYK-FTO

dependent variables

Not explicitly mentioned

control variables

Turbofect transfection protocol
HeLa cell line

positive controls

Not specified

negative controls

Not specified

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