1. Mammary cell proliferation is inhibited when LAD1 is depleted.
Mammary cell proliferation is inhibited when LAD1 is depleted. (A) MTT analysis of viability in MCF10A cells expressing shLAD1 after 4 days in culture. Data are means ± SEM of six replicates. *P < 0.05, ***P < (one-way ANOVA with Bonferroni’s correction). (B) Extracts of the indicated breast cancer lines were probed as indicated. (C) Serum-starved MDA-MB-231 cells expressing LAD1-V5 and shLAD1 MCF10A cells were stimulated with EGF (12 hours). Bromodeoxyuridine (BrdU) was added, and 60 min later, cells were stained. DAPI- and BrdU-stained nuclei were counted (in >5 fields). Data are means ± SEM of three experiments. *P < 0.05, ***P < (D) MCF10A cells (5 × 103) stably expressing shLAD1 (or shScramble) were grown in EGF-containing Matrigel (2 weeks), acini were photographed, and ImageJ was used to determine volumes. ***P < Scale bar, 100 μm. Data are from three experiments. (E) MCF10A cells were incubated with RO-3306 (10 nM; 15 hours), washed, cultured (<90 min), and stained for DAPI, LAD1 (green), and acetylated tubulin (red). (F) Cells pretreated as in (D) were fixed and probed at metaphase. Scale bar, 10 μm. (G) Cells ectopically expressing LAD1-GFP and LifeAct-cherry were pretreated as in (D). Frames taken from live cell movies are shown. Scale bar, 10 μm.
Lee Roth et al., Sci. Signal. 2018;11:eaan0949
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