Microsprayer ms ia 1c

Sixteen mice were randomly assigned to the 4 groups (N = 4). Scarb1-ASOs was dissolved in sterile saline solution and applied by a MicroSprayer MS-IA-1C (Penn-Century, Wyndmoor, PA, USA) as a single dose of 10, 40 or 100 μg in 50 μL per mouse under light anesthesia with 2.5% isoflurane. The control mice received 50 μL of saline. The volume of intratracheal administration was referred to previous report [31 (link)]. Scarb1-ASOs was administered once daily on two consecutive days. One day after the last administration, the right lung or bronchoalveolar lavage fluid (BALF) was harvested for the mRNA or protein expression analyses. The experiment with a focus on ApoB-ASOs was performed with same protocol as Scarb1-ASOs.

AF647-conjugated Scarb1-ASOs, Scarb1-ASOs, or Hprt1-ASOs were dissolved in PBS and applied by a MicroSprayer MS-IA-1C (Penn-Century, Wyndmoor, PA). AF647-conjugated Scarb1-ASOs were administered as a single dose of 0.5 µg in 50 μL of PBS. Scarb1-ASOs or Hprt1-ASOs were administered at doses of 5 μg or 20 μg per mouse in 50 μL of PBS. The control mice received 50 μL of PBS. LNA-ASOs or PBS were administered under light anesthesia with isoflurane. Two hours after the administration of AF647-conjugated Scarb1-ASOs, the lung was harvested for flow cytometry. One day after the administration of Scarb1-ASOs or Hprt1-ASOs, the bronchoalveolar cells, right lung, liver, or kidney was harvested for the mRNA expression analyses.

WT, Rac2-/- and α4Y991A mice were anesthetized with intraperitoneal injections of ketamine and xylazine (100 and 10 mg/kg body mass, respectively). Mice were administered an intratracheal instillation of 0.05 U of bleomycin sulphate (Sigma) (3 U/Kg body weight, dissolved in normal saline) or normal saline using Micro Sprayer MS-IA-1C (Penn-Century). Following the bleomycin instillation, mice were monitored daily for morbidity and mortality. Groups of WT, Rac2-/- and α4Y991A mice (n = 5 to 10 per time point) were sacrificed and their lung tissues were analyzed on day 3, 7 and 28 after bleomycin injection.