Journal: Nature communications

Article Title: Decoding the genomic landscape of chromatin-associated biomolecular condensates.

doi: 10.1038/s41467-024-51426-2

Figure Lengend Snippet: Fig. 4 | Experimental validation of potential biomolecular condensates. a Immunofluorescence images of mESC showing that SUPT6H (green) colocalizes with CTR9 (red) and SUPT5H (grey) in puncta. DNA was stained with DAPI (blue). This experimental result was consistent across two independent cell-seeding, fixation, and co-IF staining experiments (each contains three slides). Scale bar: 10 μm. b Line scans of the images of a cell co-stained for SUPT6H, CTR9 and SUPT5H, at the position depicted by the white line. The direction is from the green tick to the purple tick, and the two arrows refer to two representative puncta. FRAP experiments for SUPT6H (c), CTR9 (d) and SUPT5H (e). Left, representative images of the FRAP experiment. The white arrow refers to the punctum undergoing bleaching. Right, quantification of FRAP data for puncta of SUPT6H (n = 6), CTR9 (n = 5) and SUPT5H (n = 5). Puncta were photobleached at t = 0 s, and data were plot

Article Snippet: Following this, the ConA beads bound cells were collected using a magnet and resuspended in 100 μl of antibody buffer containing either 2 μl of DDX21 (Proteintech, 10528-1- AP, lot # 00088037), 4 μl of CTR9 (Bethyl Laboratories, A301-395A, lot # 4), 4 μl of SUPT6H (Novus Biologicals, NB100-2582, lot # 2 A), 1.5 μl of SS18 (Cell Signaling Technology, 21792 (D6I4Z), lot # 1), 1.5 μl of EP300 (Santa Cruz, sc-48343 (F-4), lot # A1323), or 0.5 μl of ELL3 (generously gifted by Prof. Chengqi Lin, Southeast University, China) primary Nature Communications | (2024) 15:6952 13 antibody respectively.

Techniques: Biomarker Discovery, Staining

Journal: Nature Chemical Biology

Article Title: Menin-MLL Inhibitors Reverse Oncogenic Activity of MLL Fusion Proteins in Leukemia

doi: 10.1038/nchembio.773

Figure Lengend Snippet: (a) MTT cell viability assay in the MLL leukemia cells KOPN-8 and MV4;11 induced by MI-2, MI-3 and MI-nc after 72h treatment. Non-MLL leukemia cell line ME-1 is shown for comparison. Data represent mean values for four samples ± s.d. Experiment was performed three times. (b) Apoptosis and cell death induced by MI-2, MI-3 and MI-nc in MV4;11 cells as detected by flow cytometry using AnnexinV/propidium iodide (PI) staining. Data represent mean values for triplicates ± s.d. (c) Selected histograms from cell cycle analysis performed by FACS after PI staining in MV4;11 cells treated with DMSO, MI-2 or MI-nc. (d) Dose-dependent effect of MI-2 on cell cycle progression measured by FACS in MV4;11 cells after PI staining, with MI-nc as a negative control. Data represent mean values for triplicates ± s.d. (e) Wright-Giemsa stained cytospins on THP-1 and MV4;11 cells after 10 days of treatment with DMSO, MI-2 or MI-nc. (f) Detection of CD11b expression in THP-1 cells assessed by flow cytometry after 6 days of treatment with DMSO, MI-2 or MI-nc. Data represent mean values for triplicates ± s.d. (g) Expression of the HOXA9 and MEIS1 genes normalized to 18S rRNA determined by qRT-PCR in THP-1 cells treated for 6 days with MI-2 and MI-3. Data represent mean values for duplicates ± s.d. Experiment was performed three times.

Article Snippet: After incubation, 1.5×10 5 cells were harvested and resuspended in 100 μl 1× Annexin V binding buffer from the Annexin V-FITC Apoptosis kit (BD Biosciences Pharmingen), incubated with 4 μl of AnnexinV-FITC and 6 μl of Propidium iodide (Sigma-Aldrich) at room temperature in the dark for 10 minutes and analyzed by flow cytometry on a LSR II instrument.

Techniques: Viability Assay, Flow Cytometry, Staining, Cell Cycle Assay, Negative Control, Expressing, Quantitative RT-PCR