Fig 2: Effect of the Bcl-6 inhibitor 79-6 on HT-29 tumor growth and microvessel densityBALB/c nude mice received subcutaneous injections of 1 × 107 tumor cells into the right flank and were subsequently treated by intraperitoneal injections of 79-6 (N = 8) or vehicle (N = 8) for 14 consecutive days. Tumor growth (E) was recorded every second day. One day after the last treatment, animals were sacrificed and cryosections of tumors were stained for the endothelial marker CD31. Analysis of microvessel density (MVD) was based on automated tissue scanning by TissueFAXS and an algorithm developed with StrataQuest 5.0 software (TissueGnostics) as exemplified in A–D. Based on fluorescence images (A) of cell nuclei (purple) and endothelial CD31 expression (green), the FITC channel (CD31) was extracted to an 8-bit image, followed by a blurring algorithm (B) after which a morphological mask (C) was applied. (D) Microvessels were identified and quantitated based on kernel radius and background threshold settings which were manually adapted for each batch of scanned images. Results (mean and standard deviation) from three independent analyses are shown in (F).

Fig 3: Mixed axonal populations of the facial nerve branches. (A) Schematic illustration of the facial nerve branches and corresponding cross-sections. The specimen is stained using anti-NF (red) and anti-ChAT (green) antibodies. Scale: 200 µm. (B) Semi-automated quantification analysis of axons in the cross-section of the mandibular branch of the facial nerve. NF-positive signals are automatically identified using the StrataQuest software (TissueGnostics, Vienna, Austria). (C) Overall and non-cholinergic axon numbers of the facial nerve branches are depicted. Data are presented as mean ± SD.

Fig 4: LRRK2 expression in colonic biopsies from PD patients and age- and gender-matched controls. (A) Original view of the represented colonic biopsy tissues. (B) All nuclei are circled with green lines, and the LRRK2-positive staining detected as positive events are circled in red lines (arrows) by the StrataQuest Analysis software version 6 (TissueGnostics, Vienna, Austria). (C) Distribution histograms of the LRRK2-positive ratio in PD patients and controls. (D) Representative pictures of LRRK2-positive cells in the submucosa of colonic samples from one 72-year-old male PD patient and one 71-year-old male control participant. Positive events are indicated by arrows. The pictures on the right are the magnification of the framed areas on the left. Scale bar: 100 µm. (E) The fractions of LRRK2-positive cells in the total cell counts in colonic biopsies were significantly higher in PD patients than controls (P = 0.004 by Mann-Whitney U-test). (F) The fractions of LRRK2-positive cells in the total cell counts in colonic biopsies from PD patients with wild type or genetic variant in LRRK2. *P < 0.05; **P < 0.01.