Fig. 4

siRNA-mediated hornerin knockdown results in altered tumor vessel parameters. a Representative whole mount Z-stacked images of CD34⁺ tumor vessels (green) from Scr siRNA (left) and Hrnr siRNA (right) 50 µm tumor sections. b Graph represents the number of CD34-positive vessels per 0.4 mm² field. Sections = 5 μm, N = 16/group (four tumors in each treatment group, four images per tumor). Vessel parameters including c vessel volume fraction, d fractal dimension, and e radius distribution in pixel units were determined from RAVE analysis of 14 Hrnr siRNA and Scr siRNAwhole-mount Z-stacks (four tumors in each treatment group, 3–4 images/tumor). f Representative images of actin, alpha-2 (ACTA2) (red) coverage on CD34⁺ tumor vessels (green) in tumor sections from Scr siRNA (left) and Hrnr siRNA (right) -treated mice. Graph represents the ratiometric quantification, as determined by dividing ACTA2-positive pixels by CD34-positive pixels, of 18 Hrnr siRNA sections (four tumors, 3–5 sections per tumor) and 20 Scr siRNA sections (four tumors, five sections per tumor). Scale bar a = 100 μm, f = 50 μm. Graphs represent mean ± SEM. *P < 0.05, **P < 0.005 by unpaired two-tailed t-test