Fig. 6

Conducting 3D-FISH validations of the PRKD3 loop in organoids of BC tissues. A Images of organoids derived from normal breast tissue (NT), primary breast cancer tissue (PT) and tamoxifen-treated breast cancer recurrent tissue (RT). Scale bar at 50 µm. B IHC staining of ER and PR in organoids of NT, PT and RT respectively with a higher magnification (40X) and a scale bar at 50 µm. C Western blotting showing ER, PR, HER2 and Ki67 protein levels in organoids of NT, PT and RT, respectively. β-actin levels were measured as a loading control. D 3D-FISH images showing interaction frequency of the PRKD3 loop in organoids of PT and RT, respectively. BAC probe combinations: promoter (green) and distal region (red) n = 50, DAPI DNA stain (blue). Square boxes in red represent the magnified view of each interaction. Scale bar at 5 µm. E Distributions of measured distances of the PRKD3 loop are significantly different for PT vs. RT (*p ≤ 0.1) by one-tail paired t test analysis. Distances were measured between the closest two foci in each nucleus