Fig 1: ACSL3 expression promotes the survival of human DTCs and BC progression. a. BC patient samples of primary tumors (n = 16), lymph nodes with solitary DTCs (n = 20) and metastasis (n = 16) were immunostained for ACSL3, Ki67 and CK. Scale bar, 500 and 50 μm. The total number of cells analyzed is 102,585 cells for primary tumors, 113,390 DTCs in lymph nodes and 70,656 cells in metastatic lesions. b. Graph showing the percentage of ACSL3high, ACSL3moderate, or ACSL3low cells in primary tumor (PT), DTCs in lymph node (LN), or mestastasis (M). ∗, P < 0.05 by Fisher's exact test. c. Graph showing the percentage of cells in PT, LN or M exhibiting either a granular cytoplasmic or perinuclear ACSL3 staining pattern. ∗∗∗∗, P < 0.0001 by Fisher's exact test. d. Kaplan–Meier plots of DFS (upper panel) or OS (lower panel) generated from 19 BC patients in this study stratified by low (n = 12) or high (n = 7) ACSL3 staining intensity of DTCs in the lymph node. Hazard ratio (HR) with 95 % confidence intervals and log-rank P value are shown. e. Representative ACSL3 (green) and TGOLN2 (red, trans-Golgi network marker) immunofluorescence of D2.0R cells seeded on BME matrices (dormant) or on BME + COL matrices (proliferative). Scale bar is 20 μm and 10 μm. f. Overlap coefficient between green (ACSL3) and red (TGOLN2) pixels (mean ± s.e.m, n = 111–118 cells per group. Comparison by Mann-Whitney test. ∗∗∗∗P ≤ 0.0001).