To construct the expression plasmids of Flag-ZFC3H1 full length (FL) or fragments, the corresponding sequences were cloned into Flag-Phage using ClonExpress® Ultra One Step Cloning Kit (Vazyme Biotech Co., Ltd). The deletion or point mutation ZFC3H1 expression plasmids were constructed by mutagenesis using the KOD-Plus-Mutagenesis Kit (Takara). ZFC3H1-FL-ZsGreen and ZFC3H1-NP-ZsGreen were constructed by deletion of the sequence between ZFC3H1 and ZsGreen in Flag-ZFC3H1-FL and Flag-ZFC3H1-NP, respectively. Plasmids encoding Flag-eIF4A3 and Flag-DDX3 were described previously (20 (link),25 (link)). To generate construct for gene editing, sgRNA targeting ZFC3H1 was synthesized, annealed and ligated to the pX330-mCherry plasmid. The sgRNA sequence is shown in Supplementary Table S1 . The Smad and β-globin constructs were described previously (28 (link)).
The CBP80, MTR4, UAP56 and ARS2 antibodies were described previously (20 (link)). Antibodies against PABPN1 (Abcam), ZFC3H1 (Novus), Flag (Sigma-Aldrich), GAPDH (Proteintech), Tubulin (Proteintech), SC35 (Sigma-Aldrich), digoxin (Roche) and SON (Thermo Fisher) were purchased. Alexa Fluor 546, Alexa Fluor 488 or Alexa Fluor 647 conjugated secondary antibodies were purchased from Life Technologies.
The CBP80, MTR4, UAP56 and ARS2 antibodies were described previously (20 (link)). Antibodies against PABPN1 (Abcam), ZFC3H1 (Novus), Flag (Sigma-Aldrich), GAPDH (Proteintech), Tubulin (Proteintech), SC35 (Sigma-Aldrich), digoxin (Roche) and SON (Thermo Fisher) were purchased. Alexa Fluor 546, Alexa Fluor 488 or Alexa Fluor 647 conjugated secondary antibodies were purchased from Life Technologies.
