INS-1 cells were seeded in a 96-well plate with further culturing for 24 h.
M2 PHARMA-May 1, 2019-LineaRx Achieves Higher Expression and Survival Rates in Human T Cells Transfected
Non-Virally with Linear DNA
The chicken macrophage (HD11) cells  were transiently transfected
with a pcDNA3-eGFP vector containing either the LILRB1R-, LILRB1S-, LILRB3R-, or LILRB3S-encoding sequence and also mock control with empty pcDNA3-eGFP vector using Lipofectamine 3000 transfection reagent (Invitrogen, USA), as per the manufacturer's recommended protocols.
The scale of GFP expression was normalized using total cell numbers transfected
with the same concentration of various constructs at the same conditions.
These cells were then transfected
with pEGS/2SS-tPA, pEGS/2SS and pEGFP-N1 vector by using LipofectamineTM 2000 Kit (Invitrogen, Shanghai, China).
Cervical cancer cell lines were transfected
with 50 nmol/L Small-interfering RNA (si-RNA) specifically targeted to NEAT1 (si- NEAT1 ) and negative control siRNA (si-NC) using lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).
GFP-LC3 plasmid in SW480 cells showed that PPAR[alpha] silence decreased autophagosome formation and GFP-LC3 puncta (Figure 1(b)), which was consistent with the transmission electron microscopy analysis (Figure 1(c)).
The cells in those seven groups were cultured in 6-well plates with 1 x [10.sup.5] cells per well, and several groups were transfected
by the corresponding adenovirus as required.
Firstly, in order to observe functions of NUCB2 in bladder cancer, T24 and 5637 cells were transfected
with specific shRNA to knockdown NUCB2.
Briefly, cells were transfected
with 100 nM siCONT, siKRAS or siHRAS, and 0.3% Lipofectamine 2000 (Life Technologies, Mulgrave, Victoria, Australia), incubated for 24 h, and removed, and fresh medium was added to wells.
The following day, cells were transfected
with mammalian expression plasmids of SLC15A3GFP or HA-SLC15A3 and control empty plasmids.