Journal: eLife

Article Title: N-terminus of Drosophila melanogaster MSL1 is critical for dosage compensation

doi: 10.7554/eLife.93241

Figure Lengend Snippet:

Article Snippet: After three rounds of washing with lysis buffer supplemented with 500 mM NaCl and TE buffer (10 mM Tris-HCl, pH 8; 1 mM EDTA), the DNA was eluted with elution buffer (50 mM Tris-HCl, pH 8.0; 1 mM EDTA, 1% SDS), treated with RNase A for 30 min and proteinase K overnight, incubated at 65 °C for 6 hr and extracted with ChIP DNA Clean&Concentrator kit (Zymo Research, USA).

Techniques: Cell Culture, Magnetic Beads, Protease Inhibitor, Sequencing, DNA Synthesis, Western Blot, Software

Journal: Molecules and Cells

Article Title: Setdb1 Is Required for Myogenic Differentiation of C2C12 Myoblast Cells via Maintenance of MyoD Expression

doi: 10.14348/molcells.2015.2291

Figure Lengend Snippet: Depletion of Setdb1 leads to downregulation of muscle-specific genes in proliferating myoblast cells. (A) Venn diagrams show overlap of genes altered by serum withdrawal, which initiates myogenic differentiation of control C2C12 cells. Depletion of Setdb1 leads to deregulation of a comparable number of genes without induction of myogenic differentiation. Only a subset of upregulated genes (51/352) in C2C12 cells with Setdb1 depletion increase during normal differentiation (left panel), while 50% of downregulated genes (207/414) overlaps with those reduced by serum withdrawal (right panel). (B) Venn diagrams show upregulated (left) and downregulated (right) genes by Setdb1 depletion that are identified by combination of oligonucleotide microarray (light gray) and RNA-seq analsyis (dark gray). (C) Setdb1 is required to maintain levels of differentiation-dependent genes. Venn diagrams show that 32 of 75 genes downregulated by Setdb1 knockdown are induced during normal myogenic differentiation (left panel), whereas only two upregulated genes overlap with genes reduced during differentiation (right panel). P-values were calculated by hypergeometric test.

Article Snippet: To better understand the underlying mechanisms linking inhibition of differentiation and depletion of Setdb1 in C2C12 myoblast cells, we investigated genes deregulated by Setdb1 depletion by applying high-density oligonucleotide microarray (Illumina MouseWG-6 v2 expression BeadChip) and direct sequencing of total RNAs ( ).

Techniques: Control, Microarray, RNA Sequencing, Knockdown