G*Power was first used to compute the required sample size. Using two-tailed t-test on previous SubQ data and high variance assumption for the tortuosity measurement, we found the required sample size to be within the range of 3 to 6. We choose to start with 6 samples (2 gel samples on each animal) per group.
All in vivo studies were conducted in compliance with the NIH Guide for Care and Use of Laboratory Animals and UCLA ARC standards. Gel implantation was performed as previously described59 (link). At day 7, the clips closing the incision were taken off and after 2 weeks, each mouse was injected with 100ul of 1mg/ml of isolectin GS-IB4-AF488 conjugate (ThermoFisher Scientific, #I21411) through the left external jugular vein before and sacrificed by isoflurane overdose. The implant hydrogels (total of 6 blank gels, 6 Fn9*10 gels, 6 Fn9(4G)10 gels) were then collected and fixed in 1% PFA for 16 hours at 4°C. Due to the variance in sample collection process, the membrane layers attached to the implants were intact only for 4 gel implant per condition. Thus, for confocal imaging, only those with intact membranes were analyzed.
All in vivo studies were conducted in compliance with the NIH Guide for Care and Use of Laboratory Animals and UCLA ARC standards. Gel implantation was performed as previously described59 (link). At day 7, the clips closing the incision were taken off and after 2 weeks, each mouse was injected with 100ul of 1mg/ml of isolectin GS-IB4-AF488 conjugate (ThermoFisher Scientific, #I21411) through the left external jugular vein before and sacrificed by isoflurane overdose. The implant hydrogels (total of 6 blank gels, 6 Fn9*10 gels, 6 Fn9(4G)10 gels) were then collected and fixed in 1% PFA for 16 hours at 4°C. Due to the variance in sample collection process, the membrane layers attached to the implants were intact only for 4 gel implant per condition. Thus, for confocal imaging, only those with intact membranes were analyzed.
