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. Author manuscript; available in PMC: 2022 Apr 6.
Published in final edited form as: Cell Rep. 2020 Oct 13;33(2):108262. doi: 10.1016/j.celrep.2020.108262

Figure 4. GIGYF1/2 Mediates Co-translational mRNA Decay.

Figure 4.

(A) Schematic representation of the DBNDD2-HA and LGALS3BP-HA CDS reporters. ORF, open reading frame; HA, hemagglutinin; STOP3, STOP positioned three codons downstream of AUG; SP, signal peptide.

(B–D) Ctrl and GIGYF1/2 KO cells were transfected with DBNDD2-HA or LGALS3BP-HA (WT or STOP3) and F-LUC-GFP. Protein samples were analyzed by western blotting using anti-HA and anti-GFP antibodies. (D) DBNDD2-HA protein levels were quantified, normalized over to those of F-LUC-GFP, and set to 100% in Ctrl cells. Bars indicate the mean value, and error bars indicate the SD (n=3).

(E–J) Cells were transfected with DBNDD2-HA (E and G) or LGALS3BP-HA (F and H) (WT or STOP3), treated with ActD, and harvested at the indicated time points.

(E–H) Show representative northern blots. (I and J) mRNA levels were quantified, as described in Figure 2A. See also Figure S2.