beta 2 Microglobulin (B2M) Human shRNA Plasmid Kit (Locus ID 567)

Snippet: hESCs were expanded and differentiated using identical protocols to hiPSC lines described above. _B2M_ KNOCKDOWN _B2M_ was knocked down using lentiviral particles containing _B2M_-targeting shRNA (OriGene, TL314543V, Virus A). Nontargeting scrambled shRNA from the same kit was used as a control. Cells were exposed to the shRNA on D14 for 18 hours at a multiplicity of infection of 8. NPCs were t

Snippet: was employed to silence the expression of HIPK1 as well as B2M as a positive control. The lenti-shRNA vectors pGFP-C-shRNA-Lenti-Hipk1, pGFP-C-shRNA-Lenti-B2M and pGFP-C-scrambled were purchased from Origene. The packaging vectors PmD2G and PsPAX.2 were obtained from Addgene. Exponentially growing 293 T cells were split and seeded at 8 × 106 cells in 100 mm dishes in RPMI 1640 medium at 3

Snippet: B2M was knocked down using lentiviral particles containing B2M-targeting shRNA (OriGene, TL314543V, Virus A). Non-targeting scramble shRNA from the same kit was used as control. Cells were exposed to the shRNA on D14 for 18 hours at MOI 8. NPCs were treated with IFNγ on D17 for 24h and fixed on D18. These experiments were carried out in biological triplicate for all three control hIPSC lines