Project description:K-GG ubiquitin profiling of SK-MEL-2 cells treated with the selective USP9X inhibitor WEHI-092 at 10 µM for 30 min, 6 h, and 24 h (and DMSO-treated control). Sample IDs:#31-35 Control 30 min SK-MEL-2#36-40 WEHI-092 30 min SK-MEL-2#41-45 Control 360 min SK-MEL-2#46-50 WEHI-092 360 min SK-MEL-2#51-55 Control 24 h SK-MEL-2#56-60 WEHI-092 24 h SK-MEL-2
Project description:K-GG ubiquitin profiling of SK-MEL-2 and UO-31 cells treated with the selective USP9X inhibitor WEHI-092 at 10 µM for 30 min, 6 h, and 24 h (and DMSO-treated control). This dataset is the matched global quantitative proteomics data.Sample IDs:#1-5 Control 30 min UO-31#6-10 WEHI-092 30 min UO-31#11-15 Control 360 min UO-31#16-20 WEHI-092 360 min UO-31#21-25 Control 24 h UO-31#26-30 WEHI-092 24 h UO-31#31-35 Control 30 min SK-MEL-2#36-40 WEHI-092 30 min SK-MEL-2#41-45 Control 360 min SK-MEL-2#46-50 WEHI-092 360 min SK-MEL-2#51-55 Control 24 h SK-MEL-2#56-60 WEHI-092 24 h SK-MEL-2
Project description:The Ewing Sarcoma cell lines A673, SK-ES-1, and SK-N-MC were treated with without 5-AZA to identify upregulated genes A673, SK-ES-1, and SK-N-MC were treated with 5-AZA
Project description:Periodontal ligament (PDL) stem-like cells (PDLSCs) are considered to be promising for the regeneration of periodontium because they demonstrate multipotency, high proliferative capacity, and potential to regenerate bone, cementum, and PDL tissue. However, the transplantation of autologous PDLSCs is restricted by limited availability and the need for tooth extraction to isolate these cells. PDLSCs are derived from neural crest cells (NCs) in early vertebrate development and NCs persist in adult PDL tissue. Therefore, we devised to promote the regeneration of periodontium by activating NCs to differentiate into PDLSCs and increasing their total number. SK-N-SH cells cultured on the extracellular matrix derived of human PDL cells (SK-PDLSCs) strongly expressed PDL-related marker genes and highly differentiated into mesenchymal lineage cells compared to untreated SK-N-SH cells. Expression levels of various hyaluronic acid (HA)-related genes were upregulated in induced pluripotent stem cells (iPSCs)-derived PDLSCs and SK-PDLSCs compared to iPSCs-derived NCs and SK-N-SH cells, respectively. Magnetic-activated cell sorting-assisted CD44-positive enrichment population of SK-N-SH cells showed higher expression of PDL-related marker genes after SK-PDLSCs induction than the CD44-negative population. Knockdown of CD44 in SK-N-SH cells significantly inhibited their ability to differentiate into SK-PDLSCs, while lower molecular HA (LHA) induction enhanced SK-PDLSC differentiation. LHA-containing electrospun nanofibrous membranes also promoted their SK-PDLSC differentiation. Our findings suggest that SK-N-SH cells are applied as a new model to induce the differentiation of NCs into PDLSCs. The LHA-CD44 relation is important for the differentiation of NCs into PDLSCs. LHA-containing electrospun nanofibrous membranes would promote the regeneration of periodontium by activating NCs in PDL tissue and increasing the total number of PDLSCs.
Project description:Ribosomes of Bacteroidia (formerly Bacteroidetes) fail to recognize Shine-Dalgarno (SD) sequences even though they harbor the anti-SD (ASD) of 16S rRNA. This is due to sequestration of the 3’ tail of 16S rRNA in a pocket formed by bS21, bS18, and bS6 on the 30S platform, which functionally occludes the ASD. Interestingly, in many Flavobacteriales, the gene encoding bS21, rpsU, contains an extended SD sequence. In this work, we present genetic and biochemical evidence that bS21 synthesis in Flavobacterium johnsoniae is autoregulated via a subpopulation of ribosomes that specifically lack bS21. Mutation or depletion of bS21 in the cell specifically increases translation of reporters with strong SD sequences, such as rpsU’-gfp. Purified ribosomes lacking bS21 (or its C-terminal region) exhibit higher rates of initiation on rpsU mRNA and lower rates of initiation on other (SD-less) mRNAs than control ribosomes. The mechanism of autoregulation depends on extensive pairing between mRNA and 16S rRNA, and exceptionally strong SD sequences, with predicted pairing free energies of < –13 kcal/mol, are characteristic of rpsU across the Bacteroidia. To our knowledge, this is the first example of biochemically-verified specialized ribosomes with a clear regulatory purpose.