Fig. 6: ENSA is linked to Stattic sensitivity in TNBC.

a Dose–response curves and half maximal inhibition concentration values of Stattic in MDA-MB-231 cells expressing control or ENSA shRNA. Dose-response curves: n = 6; Data are presented as mean ± SD. Bar plot: n = 3 independent experiments; Data are presented as mean ± SD; Two-tailed unpaired Student’s t test. b Clonogenic survival assays of MDA-MB-231 cells expressing control or ENSA shRNA and treated with 2.5 μM Stattic. n = 3. Data are presented as mean ± SD. Two-tailed unpaired Student’s t test. c Western blotting images showing the expression of pSTAT3-Tyr705, SREBP2, and ENSA in three organoids. n = 3 independent experiments. d, e Results of the cell viability assay in three TNBC patient-derived organoid models treated with 0, 5, and 15 µM Stattic. d Cell viability assay and (e) Representative bright-field images of organoids after drug treatment in three organoids. Scale bars, 200 µm. n = 3. Data are presented as mean ± SD. Two-tailed unpaired Student’s t test. f–g Stattic treatment of MDA-MB-231 cells expressing control or ENSA shRNA. Briefly, we injected shCtrl or shENSA MDA-MB-231 cells into the mammary fat pad of female NOD/SCID mice (n = 10 each). When the tumor volume reached 50–100 mm³, each group was randomly assigned to two treatment groups: vehicle and Stattic. All groups (n = 5 each) received treatment (vehicle or 10 mg/kg Stattic) three times per week after randomization. The gray arrows indicate the treatments. f The growth curve (left) and the inhibition rate of tumor volume (right). n = 5 mice per group. Data are presented as mean ± SD. Two-way ANOVA test for growth curve and two-tailed unpaired Student’s t test for inhibition rate. g The tumor weight (left) and the inhibition rate of tumor weight (right). n = 5 mice per group. Data are presented as mean ± SD. Two-tailed unpaired Student’s t test. h Scheme of the generation of the mini-PDX models for the in vivo pharmacological tests. i The relative viability of seven TNBC mini-PDX models with Stattic treatment, as normalized to vehicle treatment. n = 3 in low-ENSA group and n = 4 in high-ENSA group. Data are presented as mean ± SD. Two-tailed unpaired Student’s t test. j The relative luminance unit of each TNBC mini-PDX model treated with Stattic or vehicle. n = 5 or 6 independent capsules; Data are presented as mean ± SD. Two-tailed unpaired Student’s t test. Source data are provided as a Source Data file. n.s. not significant, Mini-PDX mini-patient-derived xenograft, RLU relative luminance unit.