Fig. 3: PINCHs drive target supramolecular assembly in cells and exhibit prolonged duration of action.
From: A pharmacological modality to sequester homomeric proteins
a, Immunostaining of U2OS cells following treatment with 250 nM BDX (left) or 250 nM EL133 (right). Keap1 is colored in magenta; DAPI staining is shown in cyan. Additional representative images are shown in Supplementary Fig. 2a. b, Quantification of Keap1 puncta in the two treatments shows a significant difference in volume (P < 0.0001, two-tailed Mann–Whitney test; n = 129 cells analyzed in two independent experiments). Bars show the mean values and a 95% confidence interval. c, Immunostaining of U2OS cells following 20 h of treatment with 2.5 μM BODIPY-conjugated PINCH EL256 and its structure. ‘R’ refers to the same structure as in Fig. 2a. Keap1 is colored in magenta; DAPI staining is shown in cyan. Additional representative images and colocalization quantification are provided in Supplementary Fig. 9. d, Lattice light-sheet microscope live imaging of U2OS cells, immediately following treatment with 2.5 μM EL256. Time after treatment is indicated in h:min. e, qPCR analysis of NQO1 expression after 20 h of treatment of OCI-AML2 cells with DMSO, EL133 or BDX at 250 nM, followed by washing of the cells. Representative results from two independent experimental replicates with two technical replicates each are shown. Bars show the mean values; error bars show the s.d. *P < 0.05, **P < 0.01 and ***P < 0.001, one-sided unpaired t-test. exp., expression.
