Fig. 6

ZFP36 regulation of SCL4A4 WT and ARE-forming mutant (Mut) reporters. a A scheme for the constructs and co-transfection experiments. b The SCL4A4 WT or the Mut 3′UTR containing Nanoluc reporters along with Firefly control plasmid were co-transfected with a Tet-O–inducible ZFP36 expression cassette (constructed as described in “Methods”) in HEK-Tet-ON cell for 16 h. ZFP36 was induced by adding 250 ng/mL of tetracycline analog, doxycycline, for additional 16 h. c Parental Hap-1 fibroblast-like cells and ZFP36 CRISPR-deleted Hap-1 cells were transfected with nanoluciferase reporters fused with WT SLC4A4 3′UTR or the Mut -SLC4A4 3′UTR along with firefly luciferase normalization vector, for 16 h. Nanoluciferase activity was measured and expressed as Nanoluc/firefly luc ratio, mean ± SEM. Statistical significance was assessed by two-way ANOVA and Student’s t-test; *P < 0.05, **P < 0.01, ***P < 0.0001 as indicated. Data are mean ± SEM of triplicate measurements of at least two independent experiments