Skip to main content
. 2016 Nov 23;6(3):518–527. doi: 10.1021/acssynbio.6b00219

Figure 1.

Figure 1

Production of a functional cyclic peptide HIF-1 inhibitor from the chromosome of human HEK-293 cells. (A) The chromosomally integrated cassette enables the conditional expression of the SICLOPPS gene, which encodes the Npu inteins with CLLFVY as the extein. Splicing of these inteins gives P1. (B) RT-qPCR of intein expression in T-REx-P1 cells incubated in normoxia or hypoxia (24 h). (C) Immunoblot of T-REx-293 and T-REx-P1 cells treated as in B. (D) Immunoblot of T-REx-HRE cells incubated with dox in hypoxia for 0–24 h. (E) P1 produced in T-REx-P1 cells has the identical HPLC retention time as the synthetic standard. (F) The mass spectrum (ESI+) of fraction shown in F from T-REx-P1 cell lysate shows the presence of P1. (G) Firefly luciferase activity in T-REx-P1 and T-REx-Scram cells transfected with TK-HRE-luciferase and incubated for under normoxia or hypoxia (16 h). Data are means (n = 3) ± SEM, **p < 0.01.