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Since this second set of cells was used for the count rate calculation, the standard deviation of this ensemble was used as an estimate of the error on the measured count rates.
One set of cells was permeabilized with 100% methanol for 10 minutes.
Immediately after the cells were combined, a set of cells was added with luciferin to obtain baseline values using the Novostar Fluorometer.
An additional set of cells was prepared which contained the NKX-RNAi knockdown construct with either the wild-type 3xFLAG-tagged NKX2.2 cDNA, or an empty vector control.
To determine protein sulfation, one set of cells was incubated with sulfate-free media (Sigma), supplemented with 500 µCi of [35S]-sulfate (PerkinElmer), with or without 100 mM sodium chlorate.
After treatment, DiI-LDL was added to each well (obtaining a final DiI concentration of 10 µg/ml protein), and for 2 h, one set of cells was incubated at 37°C and the other at 4°C.
Similar(30)
A randomly chosen set of cells is expected to have a global specialization value of one, with values >1 indicating some form of specialization.
Firstly, a set of cells is selected in order to carry out the compensation.
However, tightly controlling culture conditions whilst simultaneously monitoring the same set of cells is complicated.
A different set of cells were active when the rat was in a different area.
The cells without any treatment were served as a negative control, and another set of cells were treated with vehicle control (0.1%% dimethyl sulfoxide (DMSO)).
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