Figure 2.

TUNAR encodes a novel micropeptide, BNLN

(A) Schematic representation of BNLN reporter constructs. BNLN coding sequence (CDS) is shown in pink block. (B) Representative images (20×) and (C) activity of Renilla luciferase in HEK293T cells transfected with the indicated BNLN constructs as in (A) for 48 h. Scale bar: 50 μm. The data were normalized to firefly luciferase. ∗∗∗∗p value less than 0.0001 when compared to in-frame construct (n = 4 experimental replicates). AU, arbitrary units. (D) Representative western blots of BNLN expression in HEK293T cells transfected with the indicated BNLN constructs. CT, non-transfected cell control. (E) Representative western blots of BNLN expression in cells transfected with the indicated BNLN constructs for 48 h. (F) Levels of mRNAs encoding for BNLN in HEK293T cells expressing full-length, ATG-deleted, and CTG-deleted BNLN CDS. The data were normalized to ACTB expression level. ∗∗∗p value less than 0.0001 when compared to the full BNLN-expressing cells (n = 6, experimental replicates). Data are shown as means ± SEM; statistical significance was determined by ANOVA followed by Sidak’s multiple comparisons test in (C) and (F). p < 0.05.