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. 2015 Apr 29;3:5. doi: 10.1186/s40170-015-0131-7

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© Mattaini et al.; licensee BioMed Central. 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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N-terminal epitope-tagged PHGDH cannot support cell proliferation following PHGDH knockdown. (A) Schematic representation of the serine biosynthesis pathway. 3-PG, 3-phosphoglycerate; PHGDH, 3-phosphoglycerate dehydrogenase; NAD⁺/NADH, oxidized and reduced forms of nicotinamide adenine dinucleotide, respectively; PHP, phosphohydroxypyruvate; PSAT, phosphoserine aminotransferase; Glu, glutamate; αKG, alpha-ketoglutarate; P-Ser, phosphoserine; PSPH, phosphoserine phosphatase; Ser, serine. (B) Cell number over time of PHGDH-amplified T.T. cells stably expressing either an shRNA-resistant FLAG-His-PHGDH cDNA or empty vector (EV) control. (C) Western blot analysis assessing knockdown of endogenous PHGDH and expression of FLAG-His-PHGDH cDNA. (D) Western blot analysis of T.T. cells stably expressing an shRNA-resistant PHGDH cDNA (untagged) or empty vector (EV) control. (E) Cell number over time of the cells described in (D) when infected with virus expressing GFP or PHGDH shRNA. Error bars show standard deviation from the mean.