The transfected
cells were incubated
(5% CO2, 37 °C) for 24 h prior to imaging. Cells were
imaged in DMEM containing 25 mM HEPES (GIBCO). Live cell imaging was
performed using an inverted epi-fluorescence microscope (Axiovert135TV,
ZEISS) using a 40× oil objective. Fluorescence images were collected
by a QIClick charge-coupled device camera (QImaging). Exposure time
per fluorescence channel was 200 ms, and the images were obtained
at a frequency of 1 frame per minute for 30 to 40 min experiment duration,
depending on the logic device. Rapamycin was obtained from LClab and
GA3-AM was prepared as described previously.14 (link) Rapamycin and GA3-AM both were dissolved
in DMSO. The zero input case consisted of DMSO alone. All of these
inputs were resuspended in DMEM containing HEPES media before they
were added to the sample chamber.
cells were incubated
(5% CO2, 37 °C) for 24 h prior to imaging. Cells were
imaged in DMEM containing 25 mM HEPES (GIBCO). Live cell imaging was
performed using an inverted epi-fluorescence microscope (Axiovert135TV,
ZEISS) using a 40× oil objective. Fluorescence images were collected
by a QIClick charge-coupled device camera (QImaging). Exposure time
per fluorescence channel was 200 ms, and the images were obtained
at a frequency of 1 frame per minute for 30 to 40 min experiment duration,
depending on the logic device. Rapamycin was obtained from LClab and
GA3-AM was prepared as described previously.14 (link) Rapamycin and GA3-AM both were dissolved
in DMSO. The zero input case consisted of DMSO alone. All of these
inputs were resuspended in DMEM containing HEPES media before they
were added to the sample chamber.
