Extended Data Fig. 5: FRTAC induces a lysosomal-and FRα-dependent targets degradation.
From: Polyvalent folate receptor-targeting chimeras for degradation of membrane proteins
a, Confocal microscopy visualization and quantification of LysoTracker in HCC827 cells treated with 50 nM Atz or 10 nM FR-Atz (FAR 15) for 48 h or 1 mM LLOMe for 1 h. Scale bar, 30 μm. b, Western blot analysis of the expression levels of FRα, FRβ, EGFR or PD-L1 in different cancer cells. c, Western blot determination of the EGFR levels in SKOV3 and Hep3B cells treated with indicated concentration of FR-Ctx (FAR 15). d, Western blot analysis of the effect of FR-Ctx (FAR 15) on EGFR degradation in H292 cells transfected with siRNA against FRα or control. e, Western blot examination of the effect of FR-Ctx (FAR 15) on EGFR degradation in Hep3B cells transfected with increasing amounts of Myc-FRα (0.25, 0.5, 1 μg). f, Quantitative proteomics analysis of the changes of whole-cell H292 proteins after 48-h treatment with 10 nM FR-Atz (FAR 15) or 50 nM Atz shows no FRα degradation. Images (a) and data in b-e represent three independent experiments, data in f are the mean of three biological replicates. Data in a are shown as mean ± s.e.m. P values were calculated using an unpaired two-tailed t-tests.
