Hrp conjugated anti rabbit igg secondary antibody

To determine PfMDR1 protein levels, 20 µg whole cell protein lysates were loaded in each lane of an 8% acrylamide gel containing SDS. The proteins were transferred onto a PVDF membrane, which was then blocked O/N at 4°C with 5% milk (w/v) and 0.05% Tween-20 (ACP Chemicals, St-Leonard, QC, Canada) in phosphate buffered saline (PBS). The membrane was further incubated with the appropriate dilution of primary anti-PfMDR1 (kindly provided by Prof Cowman, Walter and Eliza Hall Institute, VIC, Australia) or anti-PfHSP70 (GenWay Biotech, San Diego, CA, USA) antibody (1:2,000) in PBS-T + 5% milk for 1 h (RT), then washed and incubated with HRP-conjugated anti-rabbit IgG secondary antibody (Abcam, Toronto, ON, Canada) (1:20,000) for 1 h (RT). Immunoreactive bands were detected with ImmunStar WesternC Chemiluminescent Kit (Bio-Rad Laboratories, Mississauga, ON, Canada) using a myECL Imager (Thermo Scientific, Burlington, ON, Canada). To confirm equal protein loading, chemiluminescence intensities of PfMDR1 were calculated for each parasite clone relative to the respective PfHSP70 chemiluminescence using ImageJ 1.47q (National Institutes of Health, USA).

Tissue samples were prepared and preserved through paraffin embedding and then dewaxed and blocked in a hydrogen peroxide/methanol solution. Antigen retrieval was performed using sodium citrate (pH 6.0) for 2×20 min at 80°C. After that, the sections were incubated with anti-SREBP2 antibody (Abcam, Cambridge, UK) and anti-IL-1β antibody (Proteintech, Chicago, USA) overnight at 4°C, followed by incubation with HRP-conjugated anti-rabbit IgG secondary antibody (Abcam) at 37°C for 30 min. The sections were stained with DAB and counterstained with hematoxylin, dehydrated in ethanol, mounted in dimethylbenzene, and placed under a coverslip. Analysis of IHC images was performed using ImageJ (NIH, Bethesda, USA).

Oridonin (99.89% purity by HPLC), SR9238 and SR‐202 were bought from MCE company. oxLDL and Dil‐oxLDL were purchased from Yiyuan biotechnology. The primary antibody against LXRα was bought from ZEN‐BIOSCIENCE. The primary antibodies against ATP‐binding cassette transporter A1 (ABCA1), ATP‐binding cassette subfamily g member 1 (ABCG1), interleukin‐1β (IL‐1β), nuclear factor NF‐kappa‐B (NF‐κB), beta‐actin and HRP‐conjugated anti‐rabbit IgG secondary antibody were bought from Abcam. The primary antibodies against FABP4 and PPARγ were bought from Proteintech Group. The GAPDH antibody and anti‐rabbit IgG Alexa Fluor® 555‐conjugated secondary antibody were obtained from Cell Signalling Technology. Dimethyl sulfoxide (DMSO) as the vehicle of oridonin was bought from MP Biomedicals. Oil Red O solution was obtained from Sigma‐Aldrich. Foetal bovine serum (FBS), DMEM medium and RPMI 1640 medium were purchased from GIBCO. Goat serum for blocking and DAPI staining solution were obtained from Solarbio.