Phosphoglucose isomerase colorimetric assay kit

The Cy5-labeled siRNAs that specifically target the EmuDdi1 gene (siEmuDdi1: CCGACTGATGGTTTCTATT) and a Cy5-labeled negative control siRNA (NC-siRNA: Cat. No. siN0000001-1-10) were designed and synthesized by RIBOBIO (Guangzhou, China). The availability of NC-siRNA was validated by searching the E. multilocularis genome (EMULTI002). Approximately 5,000 PSCs were resuspended in 200 μL Gene Pulser Electroporation Buffer (BIO-RAD) and the siRNAs were added into the buffer at a final concentration of 2.5 μM.²⁶ (link)^,27 (link) Transformations were done in a 4-mm electroporation cuvette at 200 V-100 μF using an exponential decay pulse (Bio-Rad). After incubation at 37°C for 30 min, 1mL DMEM medium was added to each electroporation cuvette and then transferred into 24-well culture plates. After 2 h of incubation, siRNA transfection efficiency was estimated with Laser Scanning Confocal Microscope (LSCM). At 6 h, 12 h, 24 h, 48 h, and 72 h post electroporation, the culture supernatants and PSCs were collected. The total protein concentration of the culture medium was examined with the BCA Protein Assay Kit (Beyotime, China). The release of PGI in the culture supernatant was measured with the Phosphoglucose Isomerase Colorimetric Assay Kit (Sigma, USA). The survival rate of PSCs based on the FDA/PI staining method was assessed by fluorescence microscopy (ZEISS).