antibody phosphorylated pdh p-pdh #31866  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Variants in glycine decarboxylase activate mechanisms of mitochondrial energy metabolism in the brain

doi: 10.1101/2025.07.12.664515

Figure Lengend Snippet: A. Representative western blots showing GLDC, GCSH and AMT in whole brain tissue from young-attenuated mutants compared to and wild type for female and male mice. B-D. Corresponding quantification of GLDC (reduced 5-6-fold), GCSH (reduced 1.5-fold) and AMT in wild type and mutant brains. E-F Representative western blots showing lipoylated DLAT in brains of young-attenuated mutants compared to and wild type for female and male mice and associated quantification. G-H. Representative western blots showing SHMT2 in brains of young-attenuated mutants compared to and wild type for female and male mice and associated quantification. I . Astrocyte model summarizing changes seen in the GCS and PDH complexes. Blue indicates reduction in GLDC, GCSH and lipoylation of DLAT. AMT is raised (red) and SHMT2 is unchanged. Astrocyte PDK4 keeps PDHE1α unchanged in a highly phosphorylated, inactivated state (black), suggesting reduction in lipoylation of DLAT may trigger alternate mitochondrial mechanisms of energy metabolism.

Article Snippet: Primary antibodies used were as follows: anti-GLDC (rabbit, Thermo Fisher, PA5-22101, 1:800), anti-Vinculin (mouse, Millipore, V9131, 1:4000), anti-AMT (rabbit, Thermo Fisher, PA5-76454, 1:3000), anti-GCSH (rabbit, ProteinTech, 16726-1-AP, 1:1000), anti-phosphorylated PDH (Ser232) (rabbit, ProteinTech, 29582-1-AP, 1:6000), anti- PDH (rabbit, ProteinTech, 18068-1-AP, 1:6000), anti-DLAT (mouse, ProteinTech, 68303-1-Ig, 1:5000), anti- α Lipoic Acid (rabbit, Abcam, ab58724, 1:2000), and anti-SHMT2 (rabbit, ProteinTech, 11099-1-AP, 1:800).

Techniques: Western Blot, Mutagenesis

Journal: bioRxiv

Article Title: Variants in glycine decarboxylase activate mechanisms of mitochondrial energy metabolism in the brain

doi: 10.1101/2025.07.12.664515

Figure Lengend Snippet: A. Representative western blots showing levels of phospho-PDHE1a and total PDHE1a levels in whole brain tissue from young-attenuated mutants (Y-GLDC ATT/ATT ) and wild type for female and male mice. B. Corresponding quantification of the ratio of p-PDHE1a/PDHE1a in wild type and mutants. C-D Model summarizing changes seen in the GCS and PDH complexes of astrocyte and neurons. C. Astrocytes . Blue indicates reduction in GLDC, GCSH and lipoylation of DLAT. AMT is raised (red) and SHMT2 is unchanged (not shown; see ). Astrocyte PDK4 keeps PDHE1a unchanged (black) in a highly phosphorylated state. Beta-oxidation of short chain fatty acids is activated (red). D. Neurons . Activation of PDH (red) detected in response to defect in astrocyte GLDC, suggests communication via the astrocyte-neuron lactate shuttle (ANLS).

Article Snippet: Primary antibodies used were as follows: anti-GLDC (rabbit, Thermo Fisher, PA5-22101, 1:800), anti-Vinculin (mouse, Millipore, V9131, 1:4000), anti-AMT (rabbit, Thermo Fisher, PA5-76454, 1:3000), anti-GCSH (rabbit, ProteinTech, 16726-1-AP, 1:1000), anti-phosphorylated PDH (Ser232) (rabbit, ProteinTech, 29582-1-AP, 1:6000), anti- PDH (rabbit, ProteinTech, 18068-1-AP, 1:6000), anti-DLAT (mouse, ProteinTech, 68303-1-Ig, 1:5000), anti- α Lipoic Acid (rabbit, Abcam, ab58724, 1:2000), and anti-SHMT2 (rabbit, ProteinTech, 11099-1-AP, 1:800).

Techniques: Western Blot, Activation Assay