Fig. 2

THRA inactivation reverses the elevated expression of ζ-globin triggered by TRIAC. A The shRNA-mediated knockdown of THRA results in a robust reduction of THRA mRNA level in K562 cells. Cells were infected with the indicated lentiviruses and then collected on the 5th day after viral infection for RNA analysis by qPCR. The lentiviral scrambled shRNA was used as the control. B THRA knockdown efficiency was assessed by Western blot using the anti-THRA antibody. β-actin: loading control. C THRA knockdown reduced the expression of ζ-globin in TRIAC-treated K562 cells. K562 stable lines expressing indicated shRNA were established and then collected after 48 h of incubation with TRIAC for RNA analysis by qPCR. D Thraa knockdown reduced the expression of ζ-globin in TRIAC-treated zebrafish embryos. Zebrafish embryos injected with the indicated MO were harvested after 24 h of TRIAC treatment for hbae5 expression by WISH at 72 hpf. hpf, hours post-fertilization. E ChIP-seq signals at the α-like globin gene cluster (K562 cells) are shown. The peaks are highlighted in green. F ChIP-qPCR assay for THRA or IgG occupancy at HS-40 in K562 cells. Data shown are the means ± SEM. Statistical significance was calculated using the Student's t test