Journal: Antioxidants

Article Title: A Multi-Target Pharmacological Correction of a Lipoyltransferase LIPT1 Gene Mutation in Patient-Derived Cellular Models

doi: 10.3390/antiox13081023

Figure Lengend Snippet: The characterization of the physiopathology of mutant LIPT1 fibroblasts. ( A ). C1 and C2: control cells. The Western blot analysis of the mutated protein LIPT1, E2 subunits of multienzyme complexes PDH and KGDH, and their lipoylated form. Actin expression and Ponceau S staining were used to demonstrate equal protein loading. ( B ). Band densitometry of Western blot data referred to actin and normalized to the mean of controls. ( C ). PDH complex activity was measured by PDH Enzyme Activity Dipstick Assay Kit. ( D ). Band intensity of PDH complex activity was obtained by ImageLab software. ( E ). KGDH activity was measured by α-Ketoglutarate Dehydrogenase Activity Assay Kit. Data represent the mean ± SD of 3 independent experiments. *** p < 0.001 and **** p < 0.0001 between the control and patient’s fibroblasts. a.u.: arbitrary units.

Article Snippet: Anti-ketoglutarate dehydrogenase subunit E2 (KGDH E2) (26865S), anti-Activating Transcription Factor 4 (ATF4) (11815S), and anti-mitochondrial transcription factor A (TFAM) (7495S) were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Mutagenesis, Control, Western Blot, Expressing, Staining, Activity Assay, Software

Journal: Antioxidants

Article Title: A Multi-Target Pharmacological Correction of a Lipoyltransferase LIPT1 Gene Mutation in Patient-Derived Cellular Models

doi: 10.3390/antiox13081023

Figure Lengend Snippet: Expression levels of LIPT1, PDH E2, KGDH E2, and their lipoylated forms in control (C1, C2, and C) and mutant (LIPT1) fibroblasts before and after the supplementation with CocT. Cells were treated with CocT for seven days. ( A ). The Western blot analysis of the mutated protein LIPT1, E2 subunits of complexes PDH and KGDH, and their lipoylated forms. Actin expression and Ponceau S staining were used to demonstrate equal protein loading. ( B ). Band densitometry of Western blot data referred to actin and was normalized to the mean of controls. ( C ). PDH complex activity was measured by PDH Enzyme Activity Dipstick Assay Kit. ( D ). Band intensity of PDH complex activity was obtained by ImageLab software. ( E ). KGDH activity was measured by α-Ketoglutarate Dehydrogenase Activity Assay Kit. Data represent the mean ± SD of 3 independent experiments. *** p < 0.001 and **** p < 0.0001 between control and mutant LIPT1 fibroblasts. ### p < 0.001 and #### p < 0.0001 between untreated and treated mutant LIPT1 fibroblasts. a.u.: arbitrary units.

Article Snippet: Anti-ketoglutarate dehydrogenase subunit E2 (KGDH E2) (26865S), anti-Activating Transcription Factor 4 (ATF4) (11815S), and anti-mitochondrial transcription factor A (TFAM) (7495S) were purchased from Cell Signaling (Danvers, MA, USA).

Techniques: Expressing, Control, Mutagenesis, Western Blot, Staining, Activity Assay, Software

Journal: Journal of Neuroinflammation

Article Title: Neuroinflammation regulates the balance between hippocampal neuron death and neurogenesis in an ex vivo model of thiamine deficiency

doi: 10.1186/s12974-022-02624-6

Figure Lengend Snippet: Neuroinflammation regulate the balance between hippocampal neuron death and neurogenesis in TD. NPC: neural progenitor cells; TD: thiamine deficiency; TPP thiamine pyrophosphate; H2A: histone 2A; H2B: histone 2B; H3: histone 3; H4: histone 4; K: lysine; R: arginine; P: phosphorylation; Ub: ubiquitination; Ac: acetylation; Me: methylation; H3K9me3: trimethylation in lysine 9 at histone 3; K3K4me3: trimethylation in lysine 4 at histone 3; TK: transketolase; PDHC: pyruvate dehydrogenase complex; Ogdh : oxoglutarate dehydrogenase; KGDHC: alfa-ketoglutarate dehydrogenase complex; PI3K/AKT: phosphatidylinositol 3-kinase/protein kinase B; BDNF: brain derived neurotrophic factor; H3K36me3: trimethylation in lysine 36 at histone 3; NSD1: nuclear receptor binding SET domain protein 1; SETD2: SET domain containing 2; Bmp4 : bone morphogenetic protein 4

Article Snippet: Rat-specific TaqMan Gene Expression Assays (Applied Biosystems) were used to detect Nsd1 (Nuclear Receptor Binding SET Domain Protein 1) (Rn01441947_m1), Bmp4 (Bone Morphogenetic Protein 4) (Rn00432087_m1), Hes5 (Hes Family BHLH Transcription Factor 5) (Rn00821207_g), Ogdh (oxoglutarate dehydrogenase) (Rn01443655_m1), Mmp9 (Matrix Metalloproteinase 9) (Rn00579162_m1), Dcx (Doublecortin) (Rn00570390_m1), Neurod1 (Neuronal Differentiation 1) (Rn01280117_m1), C3 (Complement C3) (Rn00566466_m1) and Lcn2 (Lipocalin 2) (Rn00590612_m1).

Techniques: Phospho-proteomics, Ubiquitin Proteomics, Methylation, Derivative Assay, Binding Assay