Fig 1: Contribution of the TLK2-DYNLL1 axis to the inter-transition of non-CSCs to CSCs.a Flow cytometry analysis of ALDH-positive cell rates in MDA-MB-231 cells with or without TLK2 or DYNLL1 knockdown. Means and standard deviation from biological triplicates (n = 3) are shown. (p = <0.0001; <0.0001; <0.0001; <0.0001) b Mammosphere formation assay of MDA-MB-231 cells with or without TLK2 or DYNLL1 knockdown. Scale bar, 50 μm. Means and standard deviation from biological triplicates (n = 3) are shown. (Diameter: p = <0.0001; <0.0001; <0.0001; <0.0001; Number: p = <0.0001; <0.0001; <0.0001; <0.0001) c Flow cytometry analysis of ALDH-positive cell rates in TLK2 knockdown MDA-MB-231 cells with DYNLL1 S14D or S14A mutant rescue. Means and standard deviation from biological triplicates (n = 3) are shown. (p = <0.0001; 0.1406; <0.0001) d Mammosphere formation assay in TLK2 knockdown MDA-MB-231 cells with S14D or S14A DYNLL1 mutant rescue. Scale bar, 50 μm. Means and standard deviation from biological triplicates (n = 3) are shown. (Diameter: p = <0.0001; 0.0505; <0.0001; Number: p = <0.0001; 0.0702; <0.0001) e Limiting dilution assay (LDA) in NOD/SCID mice comparing tumor formation in TLK2 knockdown versus control cells. Resected tumor xenografts were subjected to mammosphere formation assays to evaluate mammosphere-forming capacity. Means and standard deviation from biological triplicates (n = 3) are shown. (p = <0.0001) f ALDH-low (ALDHlo) MDA-MB-231 cells were sorted, re-seeded, and monitored for ALDH-positive rates over 6 days. Means and standard deviation from biological triplicates (n = 3) are shown. (p = 0.0003; 0.0011; <0.0001; 0.0004; 0.0010) g Co-culture of mNeonGreen-labeled control MDA-MB-231 cells with mRuby3-labeled TLK2 knockdown cells under 2D normal or 3D mammosphere conditions, analyzed by flow cytometry for proportional changes. Error bars represent mean ± SD (a, b, c, d, f). Statistical significance was determined by one-way ANOVA with Bonferroni correction (a, b, c, d); two-tailed unpaired Student’s t test (e, f); ns not significant; *P < 0.05; **P < 0.01; ***P < 0.001. Source data are provided as a Source Data file.