Fig. 2

A Clinical features of cases 2, 3 and 4 metastatic lesions and relative magnetic resonance imaging (case 2) or computed tomography scans (cases 3 and 4). B Representative staining results from haematoxylin–eosin (HE) and immunohistochemistry analyses of ER, PR, HER2 and Ki-67 expression in the metastatic lesions from patients 2, 3 and 4. Scale bar = 200–500 µM. ER, oestrogen receptor; PR, progesterone receptor; HER2, human epidermal growth factor. C Summary of the mutational profile of metastatic lesions of cases 2, 3 and 4. VAF refers to variant allele frequency. D Normalized enrichment score (NES) of the PI3K pathway among the four patient cases harbouring PIK3CA mutations (#1, #2, #3, #4) and the brain metastasis with an intact PIK3CA gene. All expressed genes were ranked after z score transformation, and enrichment analysis was conducted for each sample. The size of the circles reflects the percentage of genes in the core enrichment of pathway. E Representative bright-field microscopy images of organoids generated from metastatic lesions of patients 2 and 4 treated with 10 µM alpelisib for 7 days. Magnification: 10X. The number of organoids for each condition is plotted in the bar graph. F Flow cytometry of Helix NP blue-stained organoids generated from metastatic lesions of patients 2, 3 and 4 after 7 days of treatment with 10 µM alpelisib. G Number of live cells of organoids derived from metastatic lesions of patients 2 and 4 after 7 days of treatment with 10 µM alpelisib