The WEHI 231 cells were transfected with NS5A-mCherry (100 uL). A control of Thapsigargin was used, cells in this group were treated with 87.5 (0.250mM) Thapsigargin for 4 hours before RNA isolation.
T=0 cells were collected an hour after stimulation. Cells were lysed with 500uL TRIzole reagent and frozen at -80C
Cells were stimulated with 35 uL of anti-IgM 12 hours after transfection.
T=36 cells were collected and lysed with 1 mL TRIzole reagent.
RNA from both the T=0 and T=36 samples was isolated according to TRIzole manufacturing protocol.
Nanodrop concentrations were as follows:
T=0 mock 225.5 ng/uL
T= mock stim 1956.5 ng/uL
T=0 NS5A 575 ng/uL
T=0 NS5A stim 1487 ng/uL
T=0 Thap 844.5 ng/uL
T=36 mock 24322 ng/uL
T=36 mock stim 400 ng/uL
T=36 NS5A 4415 ng/uL
T=36 NS5A stim 226 ng/uL
T=36 Thap 417.5 ng/uL
The integrity of the RNA was determined using a 1% agarose gel results to follow