Fig. 2.

Evidence for translation and specific cellular localization of uPeptides in cancer-associated transcripts. a Representative immunoblots of ≥ 3 independent experiments using HEK293T cell lysates prepared 52 h after transfection of expression vectors containing indicated triple HA-tagged wt and ΔuORF TLS variants. Eight hours prior to lysis cells were exposed to proteasome inhibitor MG132. b Representative pictures of ≥ 3 independent experiments showing the expression and intracellular localization of indicated EGFP-tagged uPeptides as detected 24 h after transfection of HEK293T cells. Upstream peptides were expressed from TLS-EGFP-expression vectors containing the complete 5′-upstream sequence of indicated wt TLSs and an EGFP-tag replacing the uStop codon of the investigated uORF. The pictures shown here are presented as merged z-stack images. Additional pictures are presented in Supplementary Fig. 5