Figure 2.

The LVβ gene therapy vector containing the miR30-shRNA expression cassette can be used to reduce α-globin expression in K562 cells

(A) Schematic of α-globin mRNA showing the location of the shα1/2 and shα2 target sites. (B) Relative α1 and α2-globin mRNA expression levels in K562 cells transduced with the BB305, LVβ-shSCR, LVβ-shα1/2, and LVβ-shα2 vectors at equivalent MOIs following 5 days of hemin induction. α1/α2-globin mRNA expression levels were calculated by normalizing to β-actin in UT controls. (C) Representative western blot analysis of K562 cells transduced with the BB305, LVβ-shSCR, LVβ-shα1/2, and LVβ-shα2 vectors following hemin induction. (D and E) Relative quantification of (D) α-globin and (E) β-globin chains. The level of β-actin expression in UT cells was used to normalize signal intensity values between samples. Data represent mean ± SD of three independent experiments and vector preparations. Statistical analysis was performed using unpaired Student’s t test (∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001).