Fig. 2: rhPRG4 treatment post-injury increases blood flow and VEGF expression.

Representative images of blood flow captured through laser refraction speckle imaging, with lighter shades of blue equaling higher flow rate a, b. Quantitative analysis of blood flow data demonstrates that C57BL/6 mice treated with rhPRG4 show increased blood flow at the wound area post-injury c. The number of CD31 + blood vessels also increased in the wound area site with rhPRG4 treatment d. Immunostaining of the injury site for VEGF, CD31 and Collagen 2 demonstrated increased expression of all three proteins with rhPRG4 treatment e. VEGF levels were confirmed by western blot analysis using a tissue homogenate from the injury site, demonstrating increased VEGF expression (~39 kDa) with Histone H3 as a loading control (~18 kDa) f. Local (injury site) and systemic (serum) levels of VEGF were quantified over time post-injury by Luminex and both were found to increase with local rhPRG4 treatment g, h. Scale bar equals 2000 µm a, b; 50 µm (top panels), 10 µm (bottom panels) e. n.s. = not significant. Error bars equal mean ± SD (c, g, h 1 way ANNOVA, d – t-test). Sample sizes: a, b, c, h, n = 10 per group; d, e, f, g, n = 6 per group.