rabbit anti-phf10  (Thermo Fisher)

Journal: bioRxiv

Article Title: Differential Modulation of Polycomb-Associated Histone Marks by cBAF, pBAF, and gBAF Complexes

doi: 10.1101/2023.09.23.557848

Figure Lengend Snippet: A. schematic of the modified fire-Cas9 system showing included proximity by demerization of Frb abnd Fkpb domains by rapamycin B. Lentiviral expression constructs of fire-Cas9 components. C. Western blot showing expression of dCas9-HA and MS2-2xFrb in each cell line with wild-type TC1 negative control. D. Western blot showing expression of 2xFkbp-V5 tagged BAF subunits used for recruitment of their respective complexes. E. Co-IP pulldown with an antibody against V5 followed by Western blot showing interactions between V5 tagged BAF subunits ARID2 showing interaction with BRG1, BAF155, and PHF10-2xFkbp-V5.

Article Snippet: Protein G beads washed with PBS are incubated with each antibody, mouse α-V5 Tag (E9H8O) (1:50, Cell signaling technology), rabbit α-PHF10 (1:100, Invitrogen), and rabbit α-Arid2 (1:100, Invitrogen) in PBS for 1 hour at room temperature.

Techniques: Modification, Expressing, Construct, Western Blot, Negative Control, Co-Immunoprecipitation Assay

Journal: bioRxiv

Article Title: Differential Modulation of Polycomb-Associated Histone Marks by cBAF, pBAF, and gBAF Complexes

doi: 10.1101/2023.09.23.557848

Figure Lengend Snippet: Representative Western blot from a co-IP. Input and pulldown with an antibody against PHF10. PHF10 interacts with ARID2, BRG1, and BAF155. The PHF10 pulldown also pulls down the PHF10-2xFkbp-V5 version of the protein expressed in our pBAF FIRE-Cas9 cells, but not in wild-type TC1 cells as detected by an antibody against the V5 epitope.

Article Snippet: Protein G beads washed with PBS are incubated with each antibody, mouse α-V5 Tag (E9H8O) (1:50, Cell signaling technology), rabbit α-PHF10 (1:100, Invitrogen), and rabbit α-Arid2 (1:100, Invitrogen) in PBS for 1 hour at room temperature.

Techniques: Western Blot, Co-Immunoprecipitation Assay

Journal: bioRxiv

Article Title: Differential Modulation of Polycomb-Associated Histone Marks by cBAF, pBAF, and gBAF Complexes

doi: 10.1101/2023.09.23.557848

Figure Lengend Snippet: Chromatin immunoprecipitations against various antibodies to determine enrichment for other BAF subunits during recruitment at the Nkx2.9 locus. A. Lack of enrichment with V5 pulldowns of PHF10-2xFkbp-V5 proteins B. ARID1A enrichment with cBAF recruitments C. BAF57 enrichment with cBAF recruitments D. SS18 enrichment with gBAF recruitments E. BAF57 enrichment with pBAF recruitments. Data is presented as Rap/No Rap (recruitmenUno recruitment) normalized to the same conditions at the HoxA9 locus where no BAF complex is being recruited (n=4-6; s.e.m.)

Article Snippet: Protein G beads washed with PBS are incubated with each antibody, mouse α-V5 Tag (E9H8O) (1:50, Cell signaling technology), rabbit α-PHF10 (1:100, Invitrogen), and rabbit α-Arid2 (1:100, Invitrogen) in PBS for 1 hour at room temperature.

Techniques:

Journal: Science Advances

Article Title: ARID1A loss in neuroblastoma promotes the adrenergic-to-mesenchymal transition by regulating enhancer-mediated gene expression

doi: 10.1126/sciadv.aaz3440

Figure Lengend Snippet: ( A ) Western blot analysis of ARID1A protein expression in control (Ctrl), ARID1A +/− , ARID1A −/− #1, and ARID1A −/− #2 NGP cells. α-Tubulin was used as a loading control. ( B ) Cell growth was measured at days 1, 3, 5, and 7. Values are means ± SEM of triplicate experiments. RLU, relative light unit. ( C and D ) Transwell invasion and migration assay of the indicated cells. The fold change of the number of migrated cells passing through the membrane per field was relative to the mean for control NGP cells and was compared with the two-tailed unpaired t test (C). ** P < 0.01; *** P < 0.001. Cells were stained with 0.1% crystal violet (D). Scale bars, 100 μm. ( E ) Immunoprecipitation (IP) of the components of the BAF complexes using an anti-ARID1B antibody. PHF10 is present exclusively in the PBAF complexes and served as a negative control in this experiment; β-actin was used as a loading control. ( F ) Control luciferase shRNA (shLuc) or each of the ARID1B -targeting shRNAs (shARID1B #1 and #2) was induced by doxycycline (1.5 μg/ml) for 6 days in the indicated cells. The expression of ARID1B was detected by western blot analysis. α-Tubulin was used as a loading control. ( G ) Cell growth was measured at days 0, 3, 5, 7, and 9 after pretreatment with doxycycline (1.5 μg/ml) for 3 days to induce the expression of shLuc, shARID1B #1, or shARID1B #2. Values are means ± SEM of triplicate experiments.

Article Snippet: Proteins were detected by the anti-ARID1A rabbit polyclonal antibody (Novus Biologicals, #NB100-55334), anti-ARID1B mouse monoclonal antibody (Abcam, #ab57461), anti-BRG1 mouse monoclonal (EPNCIR111A) antibody (Abcam, #ab110641), anti-BAF155 rabbit monoclonal (D7F8S) antibody (Cell Signaling Technology, #11956), anti-SS18 rabbit monoclonal (D6I4Z) antibody (Cell Signaling Technology, #21792), anti-BAF47 mouse monoclonal (F-4) antibody (Santa Cruz Biotechnology, #sc-166164), anti-plant homeodomain finger protein 10 (PHF10) rabbit polyclonal antibody (GeneTex, #GTX116314), and anti–β-actin rabbit polyclonal antibody (Abcam, #ab8227).

Techniques: Western Blot, Expressing, Control, Migration, Membrane, Two Tailed Test, Staining, Immunoprecipitation, Negative Control, Luciferase, shRNA

Journal: Cell

Article Title: Vitamin D switches BAF complexes to protect β cells

doi: 10.1016/j.cell.2018.04.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-PHF10 , Genetex , Cat# GTX116314 RRID:AB_11176023.

Techniques: Recombinant, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software