To generate the absolute quantitative standard curve for Q-PCR analysis on lentivirus
copy number measurement, we used the PCR product of the mouse GUS gene and cloned it with the TA cloning vector (pTA[R] Easy Cloning Kit), which was purchased from Genomics BioSci and Tech (Taipei, Taiwan).
Packaging and proliferating lentivirus
with RNA interference (RNAi) technique is a conserved biological response to double-stranded RNA (Hannon, 2002), RNAi is a sequence-specific process that regulates genetic functions and provides defense against virus at the post-transcriptional level in mammalian cells and animals (Song et al., 2003; Jacque et al., 2002).
Thus, the primary receptor for EIAV, Equine Lentivirus
Receptor 1 (ELR1) was evaluated using Real Time PCR.
These cultured cortical cells at various times (from 12 to 21 DIV for different tests) were treated with memantine, AP-5, NMDA, PiB, Pin1 shRNA lentivirus
particle, control shRNA lentivirus
particle, and vehicle as indicated in the figures.
To evaluate the influence of LPS and TREM2 lentivirus
on TREM2 expression in microglia, the microglia were pretreated with LPS and/or TREM2 lentivirus
To construct stable EML cell lines expressing proinsulin, we cloned INS cDNA into the lentivirus
vector pCDH-CMV-MCSEF1-GFP-T2A-Puro, generated pseudoviral particles carrying INS, and transduced EML cells with the lentivirus
particles at an ROI of 100:1.
Suppression of HTF Proliferation by Lentivirus
Transfected VEGF shRNA.
Caption: Figure 3: miR-210 expression variation in HUVECs transduced with lentivirus
. (a) DAPI staining of HUVECs transduced with pre-210, anti-miR-210, and miR-Scr.
To produce lentivirus
, cells are transfected with packaging plasmids encoding the required gag, pol, and rev structural and regulatory genes as well as a transfer vector encoding the gene of interest (GOI).
Then, taking the purified DNA plasmids, they were transfected into human cells through lentiviral packaging to create lentivirus
. Human CD4+ T-cells were infected after the lentivirus