Journal: bioRxiv
Article Title: Pyruvate promotes ciliogenesis bypassing IFT88 dependency and attenuates DSS-induced colitis
doi: 10.64898/2025.12.17.694572
Figure Lengend Snippet: (A) Schematic illustration how pyruvate can promote tubulin acetylation. Extracellular pyruvate is transported across the plasma membrane via the monocarboxylate transporter (MCT1), then entering mitochondria through the Mitochondrial Pyruvate Carrier (MPC). In the mitochondria, pyruvate is converted to acetyl CoA which fuels the TCA cycle, citrate exits the mitochondria and is converted back to cytosolic acetyl-CoA, which serves as the acetyl donor for α-tubulin acetyltransferase (αTAT1) to acetylate α-tubulin in primary cilia microtubules. Pharmacological inhibitors targeting this mechanism are indicated: AZD3965 targets MCT1, UK5099 blocks MPC, and GM-90257 inhibits ATAT1. (B-H) CF were serum starved in basic DMEM supplemented with indicated concentrations of Pyruvate (B) Impact of pyruvate increasing concentrations (0, 1, 2, 5, 10 mM) on PC acetylation. The acetylation level was quantified using integrated density/area measurements in ImageJ. The area was delineated using Arl13b staining, and the density of acetyl tubulin staining was quantified in this area (C) Impact of pyruvate transport inhibitors AZD3965 (200nM) and UK5099 (50µM) on PC length. (D-F) Impact of sodium pyruvate or sodium acetate supplementation on PC acetylation ( D) , PC length (E) and number of PC-positive CF ( F ). (G-H) Effect of acetylation blocking using the inhibitor GM-90257 (500nM) on both PC length ( G) and number of PC-positive CF ( H) . Statistical analysis (B-H) : two-tailed unpaired t-test; *p<0.05, **p< 0.01, ***p< 0.001, ****p<0.0001.
Article Snippet: The MEK1/2 inhibitor PD98059 (used at 20μM), ERK1/2 inhibitor SCH772984 (used at 300nM), the inhibitor of pyruvate transport at the plasma membrane AZD3965 (used at 200nM), the microtubule acetylation inhibitor GM-90257 (used at 500nM) and the histone acetyl transferase inhibitors for E1A-associated Protein p300 (Ep300) C646 (used at 25μM) were purchased from MCE (MedChem Express, Monmouth Junction, NJ, USA).
Techniques: Clinical Proteomics, Membrane, Staining, Blocking Assay, Two Tailed Test
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