Affinipure goat anti mouse igg h l sa00001 1

Cisplatin (MedChemExpress, New Jersey, USA). 3-(4,5-dimetrylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Sigma-Aldrich, St. Louis, MO, USA).
The following antibodies were used: anti-Bax(50599-2-Ig), anti-Bcl-2 (12789-1-AP), anti-Cytochrome c(10993-1-AP), anti-p53(10442-1-AP), anti-HK2(22029-1-AP), anti-VDAC1(55259-1-AP), anti-β-actin(66009-1-Ig), peroxidase-conjugated AffiniPure goat anti-mouse IgG (H + L) (SA00001-1), peroxidase- conjugated AffiniPure goat anti-rabbit IgG (H + L) (SA00001-2) (Proteintech, Chicago, IL, USA); anti-G6PD (sc-373886) and anti-TIGAR (sc-166290) (Santa Cruz, CA, USA).

The cells or purified virions were lysed in hot Laemmli sample buffer and boiled for 5 min. Proteins were analyzed on a 10% SDS-PAGE gel and transferred onto a polyvinylidene difluoride (PVDF) membrane. After a 1-h block with 5% bovine serum albumin (BSA) in Tris-buffered saline-Tween, the membrane was stained with anti-RABV-G (G53, 1:2,000) or anti-RABV-N (N42, 1:1,500) overnight. Anti-RABV-G (G53) or N (N42) were prepared as previously described (Jiang et al., 2010 (link)). The membranes were then washed with Tris-buffered saline-Tween and incubated for 1 h with horseradish peroxidase-conjugated secondary antibodies (Goat pAb to Rb lgG, ab97051, Abcam, 1:4,000; HRP-conjugated Affinipure Goat Anti-Mouse IgG (H+L), SA00001-1, Proteintech, 1:4,000) at room temperature. All of the blots were developed by the west pico chemiluminescent substrate (Thermo Fisher Scientific, United States). The integrated optical density (IOD) of band signals was analyzed by the Image-Pro Plus 6 (Media Cybernetics, United Kingdom).

Cisplatin, DCA, chloroquine diphosphate salt (CQ), and 3-(4,5-dimetrylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, United States). Next, 2′,7′-dichlorofluorescin diacetate (DCFH-DA) was purchased from Beyotime Institute of Biotechnology (Shanghai, China). MitoSOX™ Red (mitochondrial superoxide indicator) and enhanced chemiluminescence (ECL) reagents were purchased from Thermo Scientific (Rockford, IL, United States). Mito-TEMPO (mitochondria-targeted superoxide dismutase mimetic) was purchased from Selleck (Houston, TX, United States). The following antibodies were used: anti-β-actin (60008-1-Ig), anti-p62 (18420-1-AP), anti-LC3B (18725-1-AP), anti-Sirt3 (10099-1-AP), anti-SOD2 (24127-1-AP), peroxidase-conjugated AffiniPure goat anti-mouse IgG (H+L) (SA00001-1), and peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H+L) (SA00001-2) (Proteintech, Chicago, IL, United States).