Journal: Cell Communication and Signaling : CCS

Article Title: PRMT5 enhances tumorigenicity and glycolysis in pancreatic cancer via the FBW7/cMyc axis

doi: 10.1186/s12964-019-0344-4

Figure Lengend Snippet: PRMT5-mediated epigenetic silencing of FBW7 leads to increased cMyc levels a. In PRMT5-silenced MIA PaCa-2 and SW1990 cells, the mRNA levels of FBW7 were increased. b. PRMT5 knockdown increased FBW7 protein levels. c. PRMT5 expression was negatively correlated with FBW7 expression in the TCGA-PAAD dataset of pancreatic cancer patients. d. In HPDE cells, overexpressing PRMT5 decreased FBW7 mRNA levels, but PRMT5 DN did not regulate FBW7 expression. e. Overexpressing PRMT5 in HPDE cells decreased FBW7 protein levels, but PRMT5 DN exerted no impact on FBW7 protein levels. f. The results of the dual luciferase assay in HEK-293 T cells showed that although PRMT5 suppressed FBW7 promoter activity, PRMT5 DN did not significantly regulate FBW7 promoter activity. g-h. The ChIP assay results demonstrated that PRMT5 occupied the promoter region enriched with CpG islands. i. PRMT5 knockdown decreased the occupancy of the heterochromatin markers H4R3me2 and H3K9me3, and the euchromatin marker H3K9ac, which reflects active transcription, was increased in MIA PaCa-2 and SW1990 cells with PRMT5 knockdown. j . Finally, we performed ChIP in HPDE cells with low PRMT5 expression. The ChIP results showed that PRMT5 increased the occupancy of heterochromatin markers such as H4MR3me2 and H3K9me3 and decreased that of the active chromatin marker H3K9ac decreased. However, the transferase-dead PRMT5 DN mutant had no such effect

Article Snippet: The anti-FBW7 IHC antibody for the detection of FBW7 was obtained from Bethyl Laboratories.

Techniques: Knockdown, Expressing, Luciferase, Activity Assay, Marker, Mutagenesis

Journal: Cell Communication and Signaling : CCS

Article Title: PRMT5 enhances tumorigenicity and glycolysis in pancreatic cancer via the FBW7/cMyc axis

doi: 10.1186/s12964-019-0344-4

Figure Lengend Snippet: PRMT5 regulates proliferation and aerobic glycolysis via the FBW7/cMyc axis a. Overexpression of wild-type FBW7 in PRMT5-overexpressing HPDE cells attenuated the increase in the cMyc protein level, but the FBW7 R465H mutant had no such effect. b. The CCK-8 assay results demonstrated that wild-type FBW7 decreased the increase in cell viability caused by PRMT5, while the FBW7 R465H mutant, which lacked enzymatic activity, did not. c. FBW7 suppressed the increase in glucose uptake caused by PRMT5 in HPDE cells, while the FBW7 R465H mutant did not. d. FBW7 inhibited the increase in lactate production induced by PRMT5 in HPDE cells, but the FBW7 R465H mutant had little impact. e-f. The ECAR measurement results showed that FBW7 mitigated the increase in glycolysis and glycolytic capacity caused by PRMT5, but the FBW7 R465H mutant did not, suggesting that PRMT5 regulates aerobic glycolysis via the FBW7/cMyc axis

Article Snippet: The anti-FBW7 IHC antibody for the detection of FBW7 was obtained from Bethyl Laboratories.

Techniques: Over Expression, Mutagenesis, CCK-8 Assay, Activity Assay

Journal: Cell Communication and Signaling : CCS

Article Title: PRMT5 enhances tumorigenicity and glycolysis in pancreatic cancer via the FBW7/cMyc axis

doi: 10.1186/s12964-019-0344-4

Figure Lengend Snippet: Schematic representation of the working model In pancreatic cancer, upregulated PRMT5 could epigenetically silence the expression of the E3 ubiquitin ligase FBW7, resulting in cMyc stabilization and a subsequent increase in the proliferation of and glycolysis in pancreatic cancer cells

Article Snippet: The anti-FBW7 IHC antibody for the detection of FBW7 was obtained from Bethyl Laboratories.

Techniques: Expressing, Ubiquitin Proteomics

Journal: eLife

Article Title: A robust method for particulate detection of a genetic tag for 3D electron microscopy

doi: 10.7554/eLife.64630

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , GTP , Chem-Impex , 00348 , .

Techniques: Transfection, Construct, Recombinant, Sequencing, In Vitro, Labeling, Electron Microscopy, Inhibition, Purification, Software