To knock-down HIF1A expression, the pGIPZ lentiviral shRNAmir that targets human HIF1A were purchased from Open Biosystems (Thermo Fisher Scientific, Inc.). We used two different shRNAs against HIF1A: V2LHS_132152 (RHS4430–98513964) (shHIF1A#A) and V2LHS_236x718 (RHS443098513880) (shHIF1A#B). A non-silencing pGIPZ lentiviral shRNAmir was used as the control (RHS4346). The production of lentivirus particles and cells infection was performed as previously described [12 (link)]. To obtain 100% GFP-positive cells, puromycin was added into the medium for an additional 10 days.
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