Supplementary material (ESI) for Chemical Communications
This journal is © The Royal Society of Chemistry 2003
Supplementary Information
Cell uptake assays with peptoids 3a-c
Cells were grown in DMEM medium supplemented with 4 mM glutamine, 10% FCS
and 100units/ml penicillin/streptomycin until 80% confluency. Cells were suspended
using trypsin/EDTA and counted. Cells were then seeded in 96 well plates at 2x104
cells per well and incubated overnight (or required time period).
Cells were washed with warm PBS buffer. Compounds 3a-3c were mixed with serum
free medium (SFM) to final concentrations of 0.1 M, 1 M, 10 M, and 20 M. To
each well 200 l of the various concentrations of 3a-3c were added and incubated at
the required temperature (4C, 20C, 37C) and for the required time period. Each
concentration was performed in triplicate. The internalisation of free fluorescein,
under the same conditions, was tested simultaneously. After incubation, cells were
washed twice with PBS and the cells analysed using fluorescence microscopy and
FACS analysis.
(a)
(b)
Figure
Analysis of cells by fluorescence microscopy
(a) HEK293T (human embryonic kidney) cells, general transmission image at low
magnification
(b) 3b (n = 5) 10M, 37ºC for 6h, fluorescent image at low magnification
The effect of NaN3 on cellular uptake with peptoids 3a-c
Cells were pre-incubated with 0.5% sodium azide in SFM for 30min. The medium
was then removed and 200l of compound 3 (10 m in SFM) in the presence and
absence of 0.5% sodium azide was added. The cells were incubated for 4 hours. The
cells were fixed and analysed by fluorescence microscopy.