Kca3.1 mice

KCa3.1-/- mice were kindly provided by Dr. James Melvin (National Institute of Dental and Craniofacial Research, Bethesda, MD, USA). As described before [16] (link), [19] (link), A group of eight-week-old male KCa3.1+/+ (C57B/6) mice (n = 8), KCa3.1-/- mice (n = 8) and eNOS-/-mice (n = 6) (Jackson laboratory, ME) weighing approximately 20 to 25 g were assigned to receive either 55 mg/kg of streptozotocin (STZ) (Sigma, MO) diluted in 0.1 M citrate buffer, pH 4.5, or citrate buffer alone by intraperitoneal injection for five consecutive days as described previously [20] (link). STZ-treated animals with blood glucose >16 mmol/l were considered diabetic. Those KCa3.1+/+ and eNOS-/-mice received citrate buffer alone served as non-diabetic controls. eNOS-/- diabetic mice were then randomized into two groups, receiving treatment with TRAM34, 120 mg/kg/day intraperitoneally or vehicle (DMSO) alone for 24 weeks. Treatment commenced within 24 h of the last STZ injection. All animals were housed in the Kearns Animal Facility of Kolling Institute of Medical Research with a stable environment maintained at 22±1°C with a 12/12-h light-dark cycle. After animals were culled, left kidneys were removed and snap frozen for the isolation of RNA or protein, and right kidneys were perfused with PBS and fixed in 10% buffered formalin for histological examination.

Wild-type (WT) C57BL/6 mice (male, 25–30 g) were procured from the Shanghai Laboratory Animal Center (Shanghai, China). KCa3.1^−/− mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA) and housed as described previously [18 (link)]. APP/PS1 transgenic mice (an AD model) were also purchased from the Jackson Laboratory (no. 003378) [15 (link)]. KCa3.1^−/− mice were crossed with APP/PS1 mice and the offspring were intercrossed to generate mice with the KCa3.1^−/−/APP/PS1 genotype. Mouse cohorts of the four genotypes (WT, KCa3.1^−/−, KCa3.1^−/−/APP/PS1, and APP/PS1) were generated and used for behavioral analysis. The protocols of all animal experiments were approved by the Animal Experimentation Ethics Committee of Shanghai Jiao Tong University School of Medicine, Shanghai, China (ethics protocol number: A-2015-010).

Male mice (10- to 12-week-old, 25–30 g) were housed in the Animal Research Center of Shanghai Jiao Tong University School of Medicine under a 12 h light/dark cycle with 23 ± 2°C and of 55 ± 5% (humidity). The experimental protocols were approved by the Animal Care and Use Committee of Shanghai Jiao Tong University School of Medicine, Shanghai, China (ethics protocol number: A-2015-010). All experiments were conducted in compliance with the ARRIVE guidelines. KCa3.1^−/− mice (Jackson Laboratory) mice have been described previously (Wei et al., 2016 (link); Yi et al., 2016a (link)). KCa3.1^−/− mice (KO) and their wildtype (WT) controls were on a C57BL/6J background.