Supplementary Source Data for: Single-molecule FRET and tracking of transfected biomolecules in living cells (https://www.cell.com/biophysj/fulltext/S0006-3495(25)00604-6; DOI: 10.1016/j.bpj.2025.09.024 ) Authors: Abhinaya Anandamurugan*, Antonia Eidloth*, Veronika Frank, Philipp Wortmann, Lukas Schrangl, Chenyang Lan, Gerhard J. Schütz, Thorsten Hugel Source_Data.zip contains the following seven data sets in h5 format. This format can be read with https://github.com/schuetzgroup/fret-analysis.git: 1) filtered-221020-fixed_esDNA-v014.tracks : Raw data for esDNA in vitro corresponding to Fig. 2b 2) filtered-241203-esDNA-R1-v014.tracks : Raw data for esDNA transfected via SLO corresponding to Fig. 2c 3) filtered-241204-esDNA-R1-v014.tracks : Raw data for esDNA transfected via SLO 4) filtered-241107-Hsp90_409_601-v014.tracks : Raw data for yHsp90_409_601 transfected via SLO corresponding to Fig. 3b+c and 4b 5) filtered-241108-Hsp90_409_601-v014.tracks : Raw data for yHsp90_409_601 transfected via SLO corresponding to Fig. 4b 6) filtered-250430-Hsp90_409_601-v014.tracks : Raw data for yHsp90_409_601 transfected via SLO corresponding to Fig. 4b 7) filtered-250508-Hsp90_409_601-v014.tracks : Raw data for yHsp90_409_601 transfected via SLO corresponding to Fig. 4b