Ab2621

1×10⁶ NUP98-NSD1 and HM-2 cells in required medium with 10% FBS and 1% Pen Strep were collected, washed, and whole cell lysate was prepared using RIPA lysis buffer supplemented with 1X proteinase inhibitor cocktail. For compound-treated samples, 1×10⁵ NUP98-NSD1 cells or 1×10⁴ HM-2 cells in required medium with 10% FBS and 1% Pen Strep were seeded in 24-well culture plates in 1 ml medium and treated with indicated doses of BT5 for 4 days. Cells were collected, counted and re-plated at initial cell density with fresh medium and compound for another 4 days. On day 8, cells were collected, washed, and whole cell lysate was prepared using RIPA lysis buffer supplemented with 1X proteinase inhibitor cocktail. Western blot detection of histone H3K36me2 (Cell signaling technology #2901S, clone CZ5H12, dilution 1:2,000), histone H3K36me3 (Cell signaling technology, #4909S, clone D5A7, dilution 1:2,000), histone H3K27me3 (Cell signaling technology, #9733S, clone C36B11, dilution 1:2,000), H3K79me3 (abcam, #ab2621, dilution 1:2,000) was performed using 12% SDS-PAGE gel. Total H3 (abcam, #ab1791, dilution 1:50,000) and Actin (Santa Cruz, #sc-47778, dilution 1:10,000 or Genescript, #A00702, dilution 1:10,000) were used as loading controls.