Fig. 2: Engineering of hNORM in mammalian cells.
From: Nitroglycerin-responsive gene switch for the on-demand production of therapeutic proteins
a, SEAP levels in culture supernatants of HEK-293 cells transfected with pMMH178 (PhPGK-sGCα-pAbGH) and pMMH179 (PhPGK-sGCβ-pAbGH) along with the reporter plasmid pCK53 (PCRE-SEAP-pAbGH). At 24 h following the transfection, the medium was changed to 100 µl of fresh medium containing either DMSO, riociguat (20 µM), DETA NONOate (100 µM) or SNAP (100 µM) for 24 h. Data are presented as mean ± s.d. (n = 3), and P values were calculated using a two-tailed, unpaired Student’s t-test. b, SEAP levels in culture supernatants of HEK-293 cells transfected with the plasmids described in the previous experiment in addition to the co-transfection of PKG1 WT (PhCMV-PKG1β-pAbGH) and/or PKG2 (PhCMV-PKG2-pAbGH). Data are presented as mean ± s.d. (n = 3), and P values were calculated using a two-tailed, unpaired Student’s t-test. c, The performance of hNORM in various mammalian cells. HEK-293, HepG2, HeLa, HDFn, CHO-K1, A549, BHK-21 and HT-1080 cells were co-transfected with pMMH178, pMMH179, PKG1β WT and pCK53. Following the transfection, DETA NONOate (100 µM) was used as an inducer for 24 h as described for the previous experiments. Data are presented as mean ± s.d. (n = 4), and P values were calculated using a two-tailed, unpaired Student’s t-test. All data shown are biological replicates. Source data are provided as a Source Data file.
