Figure 5
LSD1 is involved in breast cancer growth and is enriched in chemoresistant cells. (a) Western blots of indicated breast cancer cell line total cell extracts probed for LSD1. NPM and histone H3 were loading controls. Copped images are from samples and antibodies processed and run on the same gel. Full-length blot is shown in Supplementary Fig. 6. (b) LSD1 mRNA levels as measured by qPCR in breast cancer cell lines characterised as ER positive and ER negative. Data are expressed as arbitrary copy numbers normalised to PPIA (n = 2). LSD1 expression in: (c) local recurrent and non-recurrent breast carcinomas from GEO dataset GSE4913 (n ≥ 19); (d) primary and secondary locally recurrent breast carcinomas from GEO dataset GSE4913 (n ≥ 9); (e) Immunofluorescence microscopy was performed on matched parental and docetaxel-resistant MCF-7, T47D and MDA-MB-231 cells fixed and probed with anti-ALDH1A1, anti-PKC-θ and anti-LSD1-s111p. Graphs indicate ALDH1A, PKC- θ and LSD1-s111p TNFI and PKC and LSD1-s111p PCC (n = 20 individual cells). −1 = inverse of colocalization; 0 = no colocalization; +1 = perfect colocalization. (f) Tumour growth (mm3) up to 5 weeks post-treatment with indicated treatments (n = 5). Coloured asterisk (*) denotes significance at that time point. (g) Tumours sizes 5 weeks post-treatment from left to right: control, nab-paclitaxel 60 mg/kg, nab-paclitaxel 30 mg/kg, nab-paclitaxel 10 mg/kg, docetaxel 10 mg/kg, and docetaxel 4 mg/kg. Tumours were excised and digested into single cell suspension and then subjected to immunofluorescence microscopy where TNFI was determined for control, nab-paclitaxel 60 mg/kg, and docetaxel 10 mg/kg treated tumours after probing with: (h) anti-EGFR, anti-Snail; or (i) anti-LSD1, or anti-LSD1-s111p (n > 10). All data represents the mean ± SE. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001, Mann-Whitney test.
