Fig. 2: Specific and tunable HaloTag–azide chemistry enables intracellular pulse–chase analysis. | Communications Chemistry

Fig. 2: Specific and tunable HaloTag–azide chemistry enables intracellular pulse–chase analysis.

From: A generalizable assay for intracellular accumulation to profile cytosolic drug delivery in mammalian cells

Fig. 2

a Schematic representation of the expression of HaloTag in HT HeLa cells. WT HeLa cells are used as a vehicle. b Flow cytometry data showing HaloTag expression. Cells were treated with 50 μM TMRcl and included no TMRcl control. WT HeLa cells were used as a negative control. Data are represented as mean ± SD (n = 3). c Chemical structures of azide-tagged fluorophores. d HT HeLa cells treated with six different azido-tagged fluorophores (TMRaz, COMaz, R110az, Flaz, Cy5az, and Cy3az). Cells were first treated with 10 μM DBCOcl for 15 minutes, washed and then treated with the 50 μM azido-tagged dyes for 15 minutes. WT HeLa cells were used as a negative control. Data are represented as mean ± SD (n = 3). e SDS-PAGE analysis of cells treated with and without DBCOcl followed by TMRaz. WT HeLa cells are used as a negative control. f Fluorescent microscopy images showing “pulse-chase” experiment with cells pulsed with DBCOcl and chased with TMRaz. WT HeLa and HT HeLa cells not treated with DBCOcl were used as negative control. The scale bar represents 25 μm.

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