Journal: Cell

Article Title: METTL13 methylation of eEF1A increases translational output to promote tumorigenesis

doi: 10.1016/j.cell.2018.11.038

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Plasmid: pWZL Blast GFP , ( Orimo et al., 2005 ) , Addgene #12269.

Techniques: Virus, Plasmid Preparation, Recombinant, Protease Inhibitor, Sequencing, Modification, Bicinchoninic Acid Protein Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Silver Staining, Mutagenesis, Control, Software, Membrane

Journal: Scientific Reports

Article Title: Comparative proteomic analysis of edible bird’s nest from different origins

doi: 10.1038/s41598-023-41851-6

Figure Lengend Snippet: Quantitative analysis of EBN’s proteins. ( A ) Scatter diagram of Protein concentration ranged by iBAQ intensity from low to high. Protein was detected in all samples marked with black, and protein was detected in some samples marked with gray. ( B ) Scatter diagram of protein concentration and the difference between samples. Proteins with disulfide bonds were highlighted in blue. ( C – E ) Histogram of the iBAQ intensity of CHIA, KRT4, and PIGR in EBN samples. ( F , G ) Principal component analysis based on protein composition and quantitation. A, B, C, and D refer to Grass EBN , Imperial EBN , White EBN , and Feather EBN respectively. ( H ) Hierarchical clustering based on label-free proteome quantification. The size of the dots below refers to the iBAQ intensity of each detection. A1/A2/A3, B1/B2/B3, C1/C2/C3 and D1/D2/D3 refer to N = 3 per group.A1/A2/A3, B1/B2/B3, C1/C2/C3 and D1/D2/D3 represent Grass EBN, Imperial EBN, White EBN and Feather EBN in that order.

Article Snippet: Briefly, protein sequences were done BLAST against Gallus gallus 's reference proteome (proteome id: UP000000539_9031, with swiss-prot’s annotation) and the best match with e-value under 10 –5 was chosen as the homologous reference proteins.

Techniques: Protein Concentration, Quantitation Assay

Journal: iScience

Article Title: Increased CSN5 expression enhances the sensitivity to lenalidomide in multiple myeloma cells

doi: 10.1016/j.isci.2024.111399

Figure Lengend Snippet: Overexpression of CSN5 induces LEN sensitivity (A) Induction of LEN sensitivity in RPMI8226, KMS11, KMS12BM, and MM.1S cells overexpressing FLAG-CSN5. Empty vector (lentiEF1-FLAG-P2A-Blast) was used as a control. We used 100 μM and 1 μM LEN for LEN-resistant and -sensitive cells, respectively. Data represent the mean ± standard deviation ( n = 5). Student's t test. (B) Relationship between LEN sensitivity and the endogenous CSN5 expression levels in the 15 cell lines. Correlations between protein levels and cell viability were evaluated using simple linear regression in GraphPad Prism 8. Statistical significance was set at p < 0.05. (C) Western blots representing the expression of CUL4A and CSN5 in RPMI8226 cells overexpressing WT FLAG-CSN5 or FLAG-CSN5-H138Q. β-actin was used as a loading control. The larger proportion of neddylated CUL4A suggested that CSN5-H138Q decreased deneddylation activity. (D) LEN sensitivity in RPMI8226 cells overexpressing FLAG-CSN5-H138Q. The lentiEF1-FLAG-P2A-Blast vector harboring FLAG-tagged CSN5 or FLAG-tagged CSN5 with H138Q mutation was used for lentiviral infection with RPMI8226. We used 100 μM LEN in this assay. Data represent the mean ± standard deviation ( n = 5). Student's t test. (E) LEN response pathway (CRBN axis) involved in the antitumor mechanism of LEN. (F) Western blots of IKZF1, IKFZ3, IRF4, CSN5 expression levels after treatment with the indicated dose of LEN in FLAG-CSN5- or empty vector-overexpressing RPMI8226 cells. β-Αctin was used as the loading control. The protein bands in western blotting of IKZF1, IKZF3 and IRF4 were quantified using iBright Analysis Software. Band densities of IKZF1, IKZF3 and IRF4 were normalized to that of β-actin. Data represent the mean ( n = 2).

Article Snippet: The lentiEF1-FLAG-P2A-Blast vector was used as an Empty vector. lentiEF1-FLAG-P2A-Blast was constructed by removing the Cas9 sequence from lentiCas9-Blast (Addgene plasmid #52962; http://n2t.net/addgene:52962 ; RRID:Addgene_52962) using an In-Fusion HD Cloning Kit.

Techniques: Over Expression, Plasmid Preparation, Control, Standard Deviation, Expressing, Western Blot, Activity Assay, Mutagenesis, Infection, Software

Journal: iScience

Article Title: Increased CSN5 expression enhances the sensitivity to lenalidomide in multiple myeloma cells

doi: 10.1016/j.isci.2024.111399

Figure Lengend Snippet:

Article Snippet: The lentiEF1-FLAG-P2A-Blast vector was used as an Empty vector. lentiEF1-FLAG-P2A-Blast was constructed by removing the Cas9 sequence from lentiCas9-Blast (Addgene plasmid #52962; http://n2t.net/addgene:52962 ; RRID:Addgene_52962) using an In-Fusion HD Cloning Kit.

Techniques: Recombinant, Sequencing, Modification, Bicinchoninic Acid Protein Assay, Mass Spectrometry, Western Blot, Software