RPMI-8402 cells were plated at
10 × 106 cells per plate on 10 cm tissue culture plates
overnight before treating with DMSO (control) or 2 μMLLC0424 with 3 replicates for each treatment. After 8 h, whole
cell lysates were collected using RIPA buffer (ThermoFisher Scientific)
and quantified using Pierce Bovine Serum Albumin Standard Pre-Diluted
Set (ThermoFisher Scientific). Cell lysates were proteolyzed and labeled
with TMT 6-plex Isobaric Label Reagent (ThermoFisher Scientific) according
to the protocol from the manufacturer and subjected to 8 fractions
of liquid chromatography–mass spectrometry (LC-MS)/MS analysis
as described.33 (link) We plotted the results
using graphpad prism. To gain protein changes with high confidence,
we screened the results of (LC-MS)/MS analysis with the criteria of
peptides number >4 and unique peptides number >1.
10 × 106 cells per plate on 10 cm tissue culture plates
overnight before treating with DMSO (control) or 2 μM
cell lysates were collected using RIPA buffer (ThermoFisher Scientific)
and quantified using Pierce Bovine Serum Albumin Standard Pre-Diluted
Set (ThermoFisher Scientific). Cell lysates were proteolyzed and labeled
with TMT 6-plex Isobaric Label Reagent (ThermoFisher Scientific) according
to the protocol from the manufacturer and subjected to 8 fractions
of liquid chromatography–mass spectrometry (LC-MS)/MS analysis
as described.33 (link) We plotted the results
using graphpad prism. To gain protein changes with high confidence,
we screened the results of (LC-MS)/MS analysis with the criteria of
peptides number >4 and unique peptides number >1.
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