siRNA-mediated knockdown of TFE3

On the day before transfection, plate 100,000 cells per well in a 6 well dish.

Change media 1h 0m 0s before transfection (2mL with no antibiotics).

The next day, assemble the siRNA transfection reaction by combining 7.5µL of 20micromolar (µM) siRNA, 5µL RNAiMAX transfection reagent (Invitrogen), and 500µL Opti-MEM (Invitrogen) in a sterile 1.7ml tube.

Incubate transfection mix at Room temperature for 0h 30m 0s.

After incubation, add the transfection mixture drop wise to the cells.

Collect the lysates for immunoblotting 2 days post-transfection.