Journal: Circulation

Article Title: A Glycolytic Switch is Required for Transdifferentiation to Endothelial Lineage

doi: 10.1161/CIRCULATIONAHA.118.035741

Figure Lengend Snippet: A, ACL specific inhibitors BMS303141 and SB204990 decreased transdifferentiation yield. (n=3) B, ACL siRNA impaired transdifferentiation efficiency. (n=3) C, ACL knockdown reduced the effect of PolyI:C to induce glycolysis as reflected by ECAR measured by Seahorse assay. (n=3) D, LDHA inhibitor reduces the nuclear level of ACL induced by PolyI:C. (n=3) E. Schematic model of the control of glycolytic switch in transdifferentiation. Innate immune signaling induces a glycolytic switch and glucose is used to generate lactate or citrate, the latter is transported to the nucleus. There, it will be converted into acetyl-coA by ACL, and to support histone acetyl-coA and transdifferentiation. (*, 0.01

Article Snippet: Reagents FBPase-1 inhibitor was purchased from Alfa Aescar, (2E)-3-(3-Pyridinyl)-1-(4-pyridinyl)-2-propen-1-one (3-PO) was purchased from Calbiochem, 2-Deoxy-D-glucose (2-DG), Sodium Oxamate, Sodium dichloroacetate (DCA), Dispase I and Nile Red were purchased from Sigma, CPI613, Anacardic Acid (ANAC), BMS303141, SB204990, 8-Br-cAMP and SB431542 were purchased from Tocris Bioscience.

Techniques: Knockdown, Control

Journal: Cell Death & Disease

Article Title: Sensing soluble uric acid by Naip1-Nlrp3 platform

doi: 10.1038/s41419-021-03445-w

Figure Lengend Snippet: A Representative images of THP1 cells transduced with an empty backbone or mNaip1 at LPS-primed condition or LPS-primed and stimulated with sUA for 6 h. The membrane is in red and lipid droplets are stained for LD540, in green. B Quantification of lipid droplets per cell. C Representative images of empty backbone- and mNaip1-transduced cells primed with LPS or LPS-primed and sUA-stimulated stained with MitoTracker (red), tetramethylrhodamine ethyl ester (TMRE) (green) and DAPI (blue). The bars in each image represent 20 μm D TMRE quantification, indicating polarized mitochondria of the experiments in panel ( C ). E IL-1β Elisa of empty backbone- (red bars) and mNaip1- (blue bars) transduced cells, both at non-stimulated (Medium) condition or LPS-primed and stimulated for 6 h with sUA, citrate or palmitate. F Schematic representation of the TCA cycle and the fatty acid synthesis pathway given emphasis to the inhibitors and stimulus used in panel ( J ). G IL-1β Elisa of mNaip1-transduced and LPS-primed cells, stimulated for 6 h with sUA (200 μΜ), citrate (5 mΜ) or palmitate (100 μΜ), in the presence or absence of ATP citrate lyase inhibitor (BMS303141 at 25 μΜ), acetyl-CoA carboxylase inhibitor (TOFA at 10 μg/mL), or fatty acid synthase inhibitors (C75 at 50 μΜ or cerulenin at 5 μg/mL). In A , B , data are representative of three independent experiments and n = 12. In D , data are plotted as a median of ten different micrography fields of three independent experiments. In E , F , the experiments were performed three different times and n = 3. ** p < 0.01 and *** p < 0.001.

Article Snippet: Cerulenin (17397-89-6) was purchased from Sigma-Aldrich, and BMS303141 (CAS 943962-47-8) was purchased from Cayman Chemical.

Techniques: Transduction, Membrane, Staining, Enzyme-linked Immunosorbent Assay