Fig. 3: FRTACs mediate the lysosomal degradation of membrane proteins (EGFR, PD-L1, and CD47) via their interaction with FR. | Nature Communications

Fig. 3: FRTACs mediate the lysosomal degradation of membrane proteins (EGFR, PD-L1, and CD47) via their interaction with FR.

From: Development of folate receptor targeting chimeras for cancer selective degradation of extracellular proteins

Fig. 3

a Schematic of Ctx-FA targeting EGFR. b Dose response of EGFR degradation (24 h) in Fadu cells (n = 3). c Time course of EGFR degradation mediated by Ctx-FA (10 nM) in Fadu cells (n = 3). d Immunofluorescent staining of EGFR degradation and lysosome colocalization after treatment of Ctx (10 nM), FA (50 nM), Ctx (10 nM) + free FA (50 nM), IgG-FA (10 nM) and Ctx-FA (10 nM) for 24 h in Fadu cells. Scale bar: 25 μm. The colocalization was analyzed by Pearson’s correlation coefficients (n = 15). The intracellular fluorescence intensity is presented as mean fluorescence intensity (MFI) (n = 10). Box plot: minima (lower whisker), maxima (upper whisker), center (median), bounds of the box (25th and 75th percentiles), whiskers (range from minima to maxima). e Inhibition of EGFR degradation in the presence of Ctx-FA (10 nM) by free FA (3 mM) and Bafilomycin A1 (BAF, 200 nM) in Fadu cells for 6 h (n = 3). f Inhibition of EGFR degradation in the presence of Ctx-FA (10 nM) by MG132 (3 μM) in Fadu cells for 6 h (n = 3). g EGFR degradation mediated by Ctx-FA (10 nM) in Hela cells transfected with scramble siRNA or Rab7 siRNA for 6 h (n = 3). h Downregulation of EGFR and MAPK phosphorylation in Fadu cells. Representative blots from three biologically independent experiments. i. Schematic of Atz-FA targeting PD-L1. j, k Degradation of PD-L1 in MDA-MB-231 and A549 cells treated with Atz-FA (10 nM) for 24 h (n = 3). l Schematic of Ab2-FA targeting CD47. m, n Degradation of CD47 in MDA-MB-231 and A549 cells treated with Ab2-FA (100 nM) for 24 h (n = 3). N indicates images from three biologically independent experiments (d) or biologically independent experiments (b, c, e-g, j, k, m, n). Data are presented as mean ± SD. The statistical significance was assessed using an unpaired two-tailed t test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns: not significant. Source data are provided as a Source Data file.

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