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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Immunol Cell Biol. 2015 Jun 1;93(10):896–908. doi: 10.1038/icb.2015.59

Figure 4. hU6-induced GRB2 suppression is efficient only after removal of the full mir30 sequence.

Figure 4

(A) Left: HuT78 T cells transduced with pLKO.1 lentiviral constructs expressing hU6-driven GRB2 or LUC shRNAs flanked with or without small mir30 sequences. Right: Quantification of immunoblots. (B) Left: HuT78 T cells transduced with retro-LTR LMP or lentiviral-hU6 pLK4A constructs expressing GRB2 or LUC shRNAs flanked by the full mir30 sequence. Right: Quantification of immunoblots. Data represents 3 independent experiments with similar results. (C) Nucleotide sequence of GRB2 shRNA flanked by small mir30 sequence. (D) Nucleotide sequence of GRB2 shRNA flanked by the full mir30 sequence. (E) HuT78 T cells transduced with retroviruses carrying GRB2 or LUC microRNAs, and lentiviruses carrying hU6-GRB2 or LUC shRNAs were stimulated with 5 µg/mL anti-CD3 coated plates for 24 hrs. The supernatants were probed for IL-2 production via ELISA. The data was graphed as mean LUC secretions at 5 µg/mL anti-CD3 ±SEM from three independent experiments.