Peter van Mourik

64 Chapter 3 0 3.125 6.25 12.5 25 50 100 200 -150 0 150 300 450 [Genistein] ( µ M) DMSO VX-770 (3 µ M ) Additive 0 3.125 6.25 12.5 25 50 100 200 -500 0 500 1000 1500 2000 2500 [Genistein] ( µ M) DMSO VX-809 + VX-770 (3 µ M ) Additive a b Organoid swelling (AUC t = 60 min) Organoid swelling (AUC t = 60 min) F508del / F508del (0.05 µ M forskolin) F508del / F508del (0.128 µ M forskolin) CFTR E-cadherin C-band B-band DMSO VX-770 (10 µ M ) Genistein (50 µ M ) Curcumin (50 µ M ) DMSO CFTR E-cadherin F508del / F508del (Subject 1) F508del / S1251N F508del / F508del (Subject 2) F508del / S1251N DMSO VX-770 (10 µ M ) Genistein (50 µ M ) Curcumin (50 µ M ) DMSO C-band B-band c + VX-809 (3 µ M) + VX-809 (3 µ M) 48 h Supplementary figure S3. Dose-dependent potentiator synergy in F508del / F508del organoids. (a,b) Forskolin-induced swelling of F508del homozygous organoids stimulated with a dose range of genistein with or without VX-770 (a) or VX-770 + VX-809 (b) at the indicated concentrations of forskolin. The calculated additive re- sponses of the single treatments are indicated as a dashed line. (a and b represent data averaged from 3 F508del / F508del subjects. Each subject was measured at 2 independent culture time points in duplicate. Mean ± SD. The SD indicates the inter-subject variation). (c) Expression of CFTR and E-cadherin in whole cell lysates of F508del / F508del (2 different donors) and F508del / S1251N organoids detected by Western blot upon treatments as indicated (48 h). Of note, higher compound concentrations were used as compared to Fig. 2j,k and 3i,j. Immature (B-band) and mature (C-band) CFTR is indicated.