**2.3. Two-dimensional electrophoresis (2-DE)**

High-resolution 2-DE profiles of patatin isoforms were obtained following the procedure described in López Pedrouso et al. [10]. Briefly, total protein samples (75 μg of protein) of each biological replicate were loaded into immobilized pH gradient (IPG) strips of 24-cm long and 4–7 pH linear gradient (Bio-Rad Laboratories, Hercules, CA, USA). First dimensional isoelectric focusing (IEF) was performed in a PROTEAN IEF Cell System (Bio-Rad Laboratories) after IPG strip rehydration for 12 h at 50 V. Rapid voltage ramping was subsequently applied to reach a total of 70 kVh. Equilibration of IEF strips was performed before running second dimension using equilibration buffers. The second dimension (SDS-PAGE) was performed on 10% polyacrylamide gels using an Ettan DALTsix large vertical electrophoresis system (GE Healthcare). Second-dimension gels were run using a constant electric current of 16 mA per gel for 15 h at 25°C.
