Fig. 9.

Ribosomal stalling by non-viral sequences. (a) Scheme of the cytosolic scFv reporter constructs showing the four different C-terminal sequences of CD99L2, POTEI/POTEE, TBCA and SPIRE2 and the one from the 3′UTR of AMD1 (C-TAIL) used in place of 2A. The NPG terminal amino acids are shown in red, as well as the entire 18 aa of 2A. (b) Western blots of extracts form cells transfected with the indicated cytosolic constructs including or not a C-terminal proline. (c) Western blots of proline including constructs with POTE C-terminus or C-TAIL sequence including the addition of a C-terminal roTag as indicated. (d) Western blot of extracts of cells transfected with the secretory versions of the CD99L2 and POTE scFv constructs. Lane 1 shows the same reporter without any addition after the SV5 tag. (e) Western blots of extracts of cells transfected with the secretory reporters with CD99L2 or POTE sequences and treated, where indicated (+), with MG132 for 4 h or co-transfected, where indicated (+), with OTU or p97QQ. All panels are representative of at least n = 3 independent experiments.