Journal: Cancer research

Article Title: M 6 A RNA Methylation Regulates Histone Ubiquitination to Support Cancer Growth and Progression

doi: 10.1158/0008-5472.CAN-21-2106

Figure Lengend Snippet: ALKBH5-USP22/RNF40 axis regulates histone H2AK119 monoubiquitination and DNA damage repair. A, RNF40 has significantly enriched m6A peaks (adj P < 0.05) in ALKBH5 siRNA–transfected 143B cells compared with scrambled siRNA–transfected cells. Top two tracks (blue) represent input and bottom two show MeRIP results for scrambled siRNA and ALKBH5 siRNA–transfected 143B cells, respectively. Detected differential m6A peak sites and its fold-change values (differential peak track) were calculated by dividing total m6A value by expression level of the target site for the specific gene using MeTDiff program. B, Western blots analysis of scrambled siRNA and ALKBH5 siRNA–transfected 143B cells using antibody against RNF40. β-Actin was used as a loading control. C, qRT- PCR results showing enrichment of ALKBH5 target genes USP22 and RNF40 in 143B cells after RNA immunoprecipitation using antibodies against ALKBH5 or IgG. HPRT1 was the negative control. D, Western blots showing RNF40 and USP22 as well as and monoubiquitinated H2A (H2A ub) and H2B (H2B ub) levels in scrambled, USP22-siRNA, or RNF40-siRNA–transfected 143B cells using antibodies against the indicated proteins. β-Actin was used as the loading control. E, Immunoprecipitation assay with RNF40, CUL4A, or IgG antibody using 143B lysate. Immunoblotting is shown for RNF40 and CUL4A. F, Western blot showing RNF40, CUL4A, and DDB1 levels in scrambled or RNF40-siRNA–transfected 143B cells. β-Actin was used as a loading control. Gel picture is representative of at least three independent experiments. G, qRT-PCR results in 143B cells transfected with scrambled siRNA or ALKBH5 siRNA subjected to ChIP using antibodies against H2Aub K119. Bar graphs show histone mark enrichment near the promoter region of genes shown in the graph. Data displayed as percentages of input and error bars represent ± SEM of technical triplicates. H, FACS analysis showing GFP positive cells from DR-GFP assay reflecting DSB-induced HR repair. U2OS-DR-GFP cells were transfected with scrambled siRNA, USP22-siRNA (siUSP22), or RNF40-siRNA (siRNF40), followed by transfection with a pCAGGS vector expressing I-SceI endonuclease or empty vector as control. I, FACS showing GFP-positive cells from EJ5-GFP reporter assay reflecting total NHEJ. EJ5-GFP U2OS cells were transfected with scrambled siRNA, USP22 siRNA (siUSP22), or RNF40 siRNA (siRNF40), followed by transfection with pCAGGS vector expressing I-SceI endonuclease or empty vector as control. P values for H and I were calculated using one-way ANOVA followed by Dunnett multiple comparisons test. Bar graph shows means ± SEM (n = 3). ****, P < 0.0001.

Article Snippet: Antibodies against USP22 (#ab195289, 1:1,000) were purchased from Abcam Inc. Antibodies against P27/KIP1 (#2552), Ubiquityl-Histone H2A (Lys119) (D27C4) (#8240), Histone H2B (D2H6) (#12364), Ubiquityl-Histone H2B (Lys120) (#5546), WEE1 (#4936S), CDK2 (#2546), RNF40 (#12187S), and Cyclin B1 (#4135) were purchased from Cell Signaling Technology.

Techniques: Transfection, Expressing, Western Blot, Quantitative RT-PCR, Immunoprecipitation, Negative Control, Plasmid Preparation, Reporter Assay

Journal: iScience

Article Title: The cecropin-prophenoloxidase regulatory mechanism is a cross-species physiological function in mosquitoes

doi: 10.1016/j.isci.2022.104478

Figure Lengend Snippet:

Article Snippet: Anti-histone H2B (D2H6) antibody , Cell Signaling Technology, USA , Cat# 12364, RRID: AB_2714167.

Techniques: Recombinant, Electron Microscopy, Western Blot, Software

Journal: Cell

Article Title: Intellectual disability-associated SMARCB1 mutations reveal a nucleosome acidic patch interaction site that potentiates mSWI/SNF chromatin remodeling

doi: 10.1016/j.cell.2019.10.044

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-Histone H2B (D2H6) (WB) , Cell Signaling Technology , Cat# 12364, RRID: AB_2714167.

Techniques: Binding Assay, Purification, Recombinant, Staining, Magnetic Beads, Mutagenesis, Selection, Passaging, Bicinchoninic Acid Protein Assay, Silver Staining, Plasmid Preparation, CRISPR, Construct, Software, Imaging, Sequencing, Chromatography

Journal: Cell

Article Title: Intellectual disability-associated SMARCB1 mutations reveal a nucleosome acidic patch interaction site that potentiates mSWI/SNF chromatin remodeling

doi: 10.1016/j.cell.2019.10.044

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-Histone H2B (D2H6) (WB) , Cell Signaling Technology , Cat# 12364, RRID: AB_2714167.

Techniques: Binding Assay, Purification, Recombinant, Staining, Magnetic Beads, Mutagenesis, Selection, Passaging, Bicinchoninic Acid Protein Assay, Silver Staining, Plasmid Preparation, CRISPR, Construct, Software, Imaging, Sequencing, Chromatography

Journal: Cell metabolism

Article Title: The Mitochondrial-Encoded Peptide MOTS-c Translocates to the Nucleus to Regulate Nuclear Gene Expression in Response to Metabolic Stress

doi: 10.1016/j.cmet.2018.06.008

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-Histone H2B (D2H6) Rabbit mAb , Cell Signaling Technology , Cat# 12364; RRID: AB_2714167.

Techniques: Recombinant, SYBR Green Assay, Reporter Assay, Over Expression, Sequencing, Luciferase, Software, Real-time Polymerase Chain Reaction