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. 2017 Apr 18;36(11):1559–1576. doi: 10.15252/embj.201696392

Figure 4. Overexpression of bZIP14 activates the TCA cycle.

Figure 4

  1. Verification of bZIP14 overexpression in transformed P. tricornutum cells by qRT–PCR (n = 3). Three control (PAF6) and two bZIP14 OE lines were assessed. The control lines are empty vector transformed. Values in the y‐axis represent the relative normalized expression to the two reference genes used ± SE of three technical repeats.
  2. Determination of TCA cycle transcript levels in bZIP14 overexpression lines by qRT–PCR (n = 3). During nitrogen starvation, Urease (URE, Phatr3_J29702) transcript levels were over threefold upregulated (Dataset EV1), illustrating that the transformed cell lines did not lack nitrogen in the condition assayed here. FBA3: fructose bisphosphate aldolase 3, Phatr3_J29014; MD: malate dehydrogenase, Phatr3_J42398; FUM: fumarase, Phatr3_J36139; OGD: 2‐oxoglutarate dehydrogenase E1 component, Phatr3_J29016; CS: citrate synthase, Phatr3_J30145; SDH1: succinate dehydrogenase flavoprotein, Phatr3_J41812.
  3. Metabolite levels of the TCA cycle‐related organic acids (pyruvate, citrate, and succinate), glutamate, and gamma‐butyric acid (GABA) in bZIP14 overexpression lines (n = 6). Box plots show relative metabolite levels. The box and the horizontal line within the box represent the data at first and third quartile and median, respectively. The ends of vertical bars show maximum and minimum values. Letters indicate the results of a Tukey's test comparing metabolite levels amongst genotypes.