Figure 3
From: Assessing the Efficacy of Mdm2/Mdm4-Inhibiting Stapled Peptides Using Cellular Thermal Shift Assays
Comparisons between CETSA on whole cells and cell lysates can be used to determine drug cellular entry and target engagement.
a. The thermal stabilisation of Mdm2 by 10 μM nutlin-3 was assayed by CETSA with cell lysates and whole cells. In both cases, nutlin-3 resulted in very similar thermal shifts, as determined by the ΔΔTm50. b. The thermal shift of CETSA with whole cells with 25 μM Staplin-2 showed differences in thermal shifts. c. Confocal microscopy of HCT116 cells treated with Staplin-2FAM prior to fixation showed vesicular accumulation of the FAM-labelled peptides.
