Figure 5. Perturbing cholesterol homeostasis inhibits macrophage-mediated AR translocation.

A) PTE-82 prostate cancer cells were orthotopically injected into LysM-Cre⁺/Abca1^fl/fl/Abcg1^fl/fl or LysM-Cre⁻/Abca1^fl/fl/Abcg1^fl/fl and nuclear expression of AR was evaluated after 19 days by IHC. Representative images are shown below. n=5-6 mice per group from one of two independent experiments, data shown as the mean ± SEM. Significance was determined by unpaired t test assuming Gaussian distribution and with Welch’s correction. B) Tumor volume in mice from A, as measured by MRI. Representative T2-weighted MRI images are shown. C) Nuclear to cytoplasmic AR ratio of GFP⁺ PTE-82 cells cultured in CSS, either alone or in the presence of BMDMs derived from LysM-Cre⁺/Abca1^fl/fl/Abcg1^fl/fl or LysM-Cre⁻/Abca1^fl/fl/Abcg1^fl/fl donors. 6-8 randomly selected images from two wells of the chamber slides were pooled for analysis. Data reflects one of three independent experiments. Significance was determined by Mann-Whitney. D) Expression of Nr1h2 (LXRβ) and Nr1h3 (LXRα) in prostate cancer cell lines and BMDMs. n=3, data shown as the mean ± SEM from one of two independent experiments. E) Nuclear to cytoplasmic AR ratio of GFP⁺ PTE-82 cells cultured in CSS or co-cultured with BMDMs. 5 μM SR9243 (LXRα/β inverse agonist) or 5 μM RGX-104 (LXRβ agonist) were added to co-cultures as indicated. 6-8 randomly selected images from two wells of the chamber slides were pooled for analysis. Data reflects one of three independent experiments. Significance determined by Mann-Whitney.