Quantity 1 software

Number and proportion of specifically stained positive or negative cells were counted from cell cultures, or they were quantified from a series of sequentially representative cut tissue sections. Single PNs in cerebellar cell cultures and in the cerebellar cortex were randomly selected and photographed by confocal microscopy with a ×63 oil immersion objective. Quantity I software (Bio-Rad) was used for the quantitative analyses of cDNA and protein bands on gel or film. Comparison between either WT and pcd groups or experimental and control groups was performed with 2-tailed Student’s t test or 1-way ANOVA. Results were listed as mean ± SD. Differences were rated significant when P values were less than 0.05 or very significant when P values were less than 0.01.

Denaturing SDS-polyacrylamide gel electrophoresis and Western blotting was performed as described elsewhere [42 (link)]. Dilutions of 1:4000 for DENV NS3 monoclonal antibody (GeneTex International Corporation, Hsinchu City, Taiwan), 1:500 for human HIF-1α mouse monoclonal antibody (kindly provided by G. Simos, originally obtained by BD Biosciences, San Jose, CA, USA), 1:1000 for human phosphorylated AKT rabbit monoclonal antibody (Ser⁴⁷³, Cell Signaling, Leiden, The Netherlands), 1:100 for GFP rabbit polyclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA), and 1:6000 for β-actin mouse monoclonal antibody (Merck-Millipore, Burlington, MA, USA), respectively, were used. A dilution of 1:2000 for the secondary anti-mouse and anti-rabbit horseradish peroxidase-conjugated antibodies (Cell Signalling, Leiden, The Netherlands) was used. Imaging quantification was performed by using Quantity I software (Bio-Rad, Hercules, CA, USA).