Fig. 3: Silencing of MMP9 by MMP9 siRNA loaded nanoparticles worsened CAC conditions in mice.

a (i) Immunofluorescence staining of nanoparticles loaded with FITC fluorescein dye as a probe after 12 and 24 h of delivery is shown by red arrows. DAPI was used to counterstain nuclei with blue fluorescence. Scale bars: 50 mm. Images were taken with ×20 magnification, and (ii) WB of the whole-cell lysates (25 μg/lane) extracted from mucosal stripping of WT mice (n = 5 per group, two individual experiments) with and without CAC gavaged with either MMP9 siRNA or scrambled siRNA loaded nanoparticles probed with anti-MMP9. GAPDH was used as the loading control for the blots. Bar graph presentation of the b body weight gain (in percent), c number of dysplastic lesions in the whole colon assessed under microscope, and d number of polyps in the whole colon assessed under microscope among WT mice (n = 5 per group) with and without CAC gavaged with either MMP9 siRNA or scrambled siRNA loaded nanoparticles. Each bar represents mean ± S.E., **p < 0.005 and *p < 0.05. e H&E staining of the colonic Swiss rolls of WT mice (n = 4 per group) with and without CAC gavaged with either MMP9 siRNA or scrambled siRNA loaded nanoparticles. Images were taken with ×10 and ×20 magnifications. Black arrows indicate flat polyp and blue arrows indicate dysplasia in epithelium respectively. Scale bars: 50 mm. f Bar graph presentation of the histological score calculated on three parameters- infiltration of neutrophils, loss of crypt architecture, and foci of ulceration. Each bar represents mean ± S.E., *p < 0.05.