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. 2024 Jun 10;13:RP93094. doi: 10.7554/eLife.93094

Figure 4. Effect of 7,8-dihydroxyflavone (7,8-DHF) on the pyridoxal 5’-phosphate (PLP)/PL ratio in cultured hippocampal neurons from WT or knockout of pyridoxal 5’-phosphate phosphatase (PDXP-KO) mice.

Figure 4.

(a) Effect of long-term PDXP deficiency on total PLP levels in hippocampal neurons. Data are mean values ± SE of n=4 biologically independent experiments. Statistical significance was assessed with a two-tailed, unpaired t-test. A representative image of primary hippocampal neurons stained for the neuronal marker protein MAP2 is shown in the insert (pixel intensities were color-inverted for better visualization). Scale bar, 100 µm. (b) Western blot analysis of PDXP and pyridoxal kinase (PDXK) expression in hippocampal neuron samples shown in (a). The same blots were reprobed with α-actin antibodies as a loading control. The densitometric quantification of PDXK signals is shown on the right; data are mean values ± SE of n=4 biologically independent experiments. (c) Effect of 7,8-DHF (20 µM, 45 min) or the DMSO solvent control (0.02% vol/vol, 45 min) on the PLP/PL ratio in hippocampal neurons of PDXP-WT or PDXP-KO mice. Source data are available for this figure.

Figure 4—source data 1. Quantification of pyridoxal 5’-phosphate (PLP) and PLP/PL levels in hippocampal neurons (to Figure 4a and c).
Data are from high-performance liquid chromatography (HPLC)-based measurements.
Figure 4—source data 2. Quantification of western blots (to Figure 4b).
Densitometric quantification of pyridoxal kinase (PDXK) and actin levels in hippocampal neurons derived from pyridoxal 5’-phosphate phosphatase (PDXP)-WT and knockout of PDXP (PDXP-KO) mice.
elife-93094-fig4-data2.xlsx (451.6KB, xlsx)