Extended Data Fig. 11: YAP inhibition suppresses post-chemotherapy regrowth.
From: Cell–matrix interface regulates dormancy in human colon cancer stem cells
a–c. CCO32 organoids were cultured in the indicated conditions with or without CPT (10 nM) and TEADi (20 μM) (left). Organoid area per well (mean ± s.e.m.) are shown on the right (a, b). Total DAPI-positive area to total organoid area (mean ± s.e.m.) is shown (c). two-sided Welch’s t-test. d. The constructs of the Dox-inducible shYAP/TAZ cassettes. e. Knockdown efficiencies of YAP1 (encoding YAP) and WWTR1 (encoding TAZ) in the indicated CCO lines. shYAP/TAZ lentivirus-infected organoids were cultured with or without 500 ng/ml Dox for 2 days. Data show mean values. f. The expression of YAP/TAZ/TEAD target genes (LATS2, CTGF and CYR61) (mean ± s.e.m.) relative to the control in shYAP/TAZ lentivirus-infected CCO32 organoids. Organoids were cultured with or without 500 ng/ml Dox for 2 days. From the left, p = 5.7 x 10–07, p = 7.5 x 10–05, two-sided Welch's t-test. g. The growth of shYAP/TAZ CCO7, CCO20 and CCO32 organoids treated with or without 500 ng/ml Dox (left). The fluorescence of mRFP1 in the shTAZ construct was captured. l. Colony formation efficiency of Dox-treated (500 ng/ml) shYAP/TAZ organoids relative to the untreated control (right). Each dot shows one well. Statistics, two-sided Welch’s t-test. h. Representative images demonstrating the regrowth of YAP/TAZ-knockdown CCO7, CCO20 and CCO32 organoids following CPT treatment (CCO7; 50nM, 24 hr, CCO20 and CCO32; 10 nM, 24 hr) with or without Dox (500 ng/ml) (left). The total area of organoids per well (mean ± s.e.m.) in the indicated conditions (right). For each condition, 1,000 cells were plated, and the images were captured on day 7 (CCO7, CCO32) or day 10 (CCO20) after CPT treatment. Each dot shows one well. Statistics, two-sided Welch’s t-test. Scale bars: 50 μm (c), 1 mm (a, k)
