Fig. 1
The i.p. injection improves the delivery of AAV2/8 CAV1 to the mouse gallbladder compared with i.v. injection
Mice were injected (either via the i.v. or i.p. route) with or without AAV2/8CAV1 at 1 × 1011 vg/animal and then assigned to chow or LD (8-week)
The letters on each bar are provided for statistical purposes, and different letters indicate significance (P < 0.05). If there are no significant differences between two bars, they have the same letter. The individual P values are described in Supplementary Table 3.
A. Serial sections of frozen (5 μm) liver, gallbladder, and ileum tissues collected from mice were stained for β-actin (red) to quantify cell-specific CAV1 (green) transduction percentages. Representative images are shown. Blue: DAPI staining for nuclei. Scale bar: 20 μm. Quantification of liver, gallbladder, and ileum transduction was determined by the percentage of CAV1 staining-positive area. The fluorescence intensities of CAV1 expression areas were normalized to the fluorescence intensities of the β-actin-stained area. The results in the lower panel are shown as the mean ± SD (three sections from each animal were analyzed). n = 13 for each group, mice with either i.v. or i.p. administration of AAV2/8CAV1; n = 5 for control (uninjected) mice.
B. qRT‒PCR analysis of AAV genomes in liver, gallbladder, and ileum collected from chow-fed (n = 9 each group, mice with either i.v. or i.p. administration of AAV2/8CAV1) or LD-fed (n = 13 for each group, mice with either i.v. or i.p. administration of AAV2/8CAV1) mice.
C. Cryosections (5 μm) of liver and gallbladder tissues collected from mice were stained for β-actin (red) to quantify cell-specific CAV1 (green) transduction percentages. Representative images are shown. Blue: DAPI staining for nuclei. Scale bar: 100 μm. GB, gallbladder.
D. Quantification of liver and gallbladder transduction was determined by the percentage of CAV1 staining-positive area. The fluorescence intensities of CAV1 expression areas were normalized to the fluorescence intensities of β-actin-stained areas under the same fluorescence microscopic field. The results in the right panel are shown as the mean ± SD (three sections from each animal were analyzed). n = 13 for each group, mice with either i.v. or i.p. administration of AAV2/8CAV1; n = 5 for control (uninjected) mice. *, P = 0.047 for the fluorescence intensities of CAV1 expression areas in liver versus GB in chow-fed control mice; #, P = 5.7e-14 for the fluorescence intensities of CAV1 expression areas in liver versus GB in chow-fed i.v. AAV2/8CAV1-injected mice; ▲, P = 5.95e-13 for the fluorescence intensities of CAV1 expression areas in liver versus GB in LD-fed i.v. AAV2/8CAV1-injected mice.
E. Western blotting analysis of CAV1 protein expression in the liver, gallbladder, and ileum of chow-fed or LD-fed mice. β-actin was used as a loading control. The data are representative of three samples for each protein. i.v., AAV2/8CAV1 injection (1 × 1011 vg/mice) via the i.v. route; i.p., AAV2/8CAV1 injection (1 × 1011 vg/mice) via the i.p. route.
F. Western blotting analysis for the comparison of CAV1 protein expression in the gallbladder of chow-fed or LD-fed mice between i.v. (1 × 1011 vg/mouse) and i.p. (1 × 1011 vg/mouse) routes of administration of AAV2/8CAV1. β-actin was used as a loading control. The data are representative of three samples for each protein.