Skip to main content
. 2017 Aug 2;37(4):BSR20170919. doi: 10.1042/BSR20170919

Figure 4. Validate EGFR SUMO 2/3 modification.

Figure 4

(A) A431 cells were harvested with BlastR lysis buffer with or without NEM. Lysates were incubated with ASM24 beads. Samples were separated by SDS/PAGE and analyzed by Western blot for total SUMOylated proteins with SUMOylated 2/3-HRP antibody. (B) Untreated and 15 min EGF-treated lysates were incubated with ASM24 beads. IgG control beads were incubated with untreated A431 lysate with NEM to identify non-specific binding. Samples were separated by SDS/PAGE and analyzed by Western blot for SUMOylated 2/3 EGFR with an EGFR antibody. (C) Untreated and 15 min EGF-treated lysates with or without NEM were incubated with EGFR antibody and protein G beads. Protein G beads alone were added to untreated A431 lysate with NEM to identify non-specific bead binding. Samples were separated by SDS/PAGE and analyzed by Western blot for SUMOylated 2/3 EGFR with sumoylated 2/3-HRP antibody. (D) Untreated and 15 min EGF-treated lysates with or without NEM were incubated with EGFR antibody and protein G beads. Protein G beads alone were added to untreated A431 lysate with NEM to identify non-specific bead binding. Samples were separated by SDS/PAGE and analyzed by Western blot for total EGFR with an EGFR antibody.