Erratum for "Constitutively active Rheb mutants [T23M] and [E40K] drive increased production and secretion of recombinant protein in Chinese hamster ovary cells" (Biotechnology and Bioengineering, (2021), 118, 7, (2422-2434), 10.1002/bit.27748)

This erratum is being published to correct the versions of Figures 3 and 5. On page 2428, Figure 3 has been revised, the differences are in terms of some of the images (immunoblots) shown in panels (e) and (f). On page 2431, Figure 5 has been revised, the earlier version of this figure contained an incorrect version of panel (g) and did not contain the current panel (h). The revised versions of the Figures 3 and 5 are shown below. 3 Figure (Figure presented.) Rheb mutants drive constitutive mTORC1 signaling in GLuc-CHO cells. (a) SDS-PAGE and western blot analysis of lysates harvested from GLuc-CHO cells transiently expressing the indicated Rheb mutants or transfected with an empty vector (EV); some were also cotransfected with FLAG-TSC1/2, as indicated. The growth medium was replaced with medium lacking FBS for 16 h followed by D-PBS for 60 min immediately before harvesting. (b) As in (a) for FLuc-CHO cells. (c) GLuc-CHO cells expressing the indicated Rheb mutants or an empty vector (EV) were grown in fully supplemented media with cell number recorded every 24 h for 7 days. (d) As in (c) with cells grown in media lacking FBS. Figures are representative of mean ± SD for three independent experiments. (e) BrdU incorporation assay for FLuc-CHO cells expressing the indicated Rheb mutants or transfected with EV. Cells were allowed to grow in fully supplemented medium for 24 h before addition of BrdU for 1 h. (f) As in (e) for GLuc-CHO cells. All graphs represent mean ± SD for three independent experiments. The data were analyzed by Student's t-test compared to EV. *p < 0.05; ***p < 0.001. D-PBS, Dulbecco's-phosphate-buffered saline; FBS, fetal bovine serum; GLuc-CHO, Gaussia Luciferase-Chinese Hamster Ovary; mTORC1, mechanistic target of rapamycin complex 1; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SD, standard deviation 5 Figure (Figure presented.) Rheb T23M and E40K drive increased secretion of mAb from CHO-S cells. (a) SDS-PAGE western blot analysis of lysates harvested from ExpiCHO cells stably overexpressing the indicated Rheb mutant or endogenous Rheb (Endo) (b) SDS-PAGE western blot analysis of lysates harvested from ExpiCHO cells stably overexpressing the indicated Rheb mutant or transfected with empty vector (EV). (c) ExpiCHO cells stably expressing the indicated Rheb mutant or only the endogenous Rheb (Endo) were grown for 48 h before treatment with 30 µM puromycin for 30 min. Lysates were then harvested for SuNSET assays. (d) ExpiCHO cells stably expressing the indicated Rheb mutants or only the endogenous Rheb (Endo) were allowed to proliferate with cell number recorded every 24 h for 10 days. (e) BrdU incorporation assay of ExpiCHO cells stably expressing the indicated Rheb mutants or only endogenous Rheb (Endo). Cells were allowed to proliferate for 24 h before addition of BrdU for a further 1 h. (f) Transient IgG yield from ExpiCHO cells stably expressing the indicated Rheb mutant or endogenous Rheb after 10 days of expression. (g) Specific productivity (Qp) of ExpiCHO cells stably expressing the indicated Rheb mutant. Figure represents n = 1. (h) SDS-PAGE western blot analysis of cell lysates from (g). All figures other than (g) are representative of three independent experiments. All graphs represent mean ± SD for three independent experiments. The data were analyzed by Student's t-test compared with Endo. *p < 0.05; **p < 0.01; ***p < 0.001. CHO, Chinese Hamster Ovary; SD, standard deviation; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SuNSET, surface sensing of translation All other parts are unchanged. The authors regret these errors. The published article can be found here: https://onlinelibrary.wiley.com/doi/full/10.1002/bit.27748.