pGCTB-SCs were seeded in 6-well dishes at a 1.2 × 106 cells/well density and incubated overnight. The following day, the culture media were replaced for each reagent, and the cells were incubated for an additional 12 h. After incubation, the cells were washed twice with ice-cold PBS, scraped, and centrifuged. Cytoplasmic and nuclear proteins were isolated using nuclear and cytoplasmic extraction reagents (Thermo Fisher Scientific), to which Cell Lytic M (Sigma-Aldrich) with protease inhibitor cocktail (cOMplete™ Mini: Sigma-Aldrich) were added.
Western blotting was performed as previously described 45 (link) with the following primary antibodies: β-catenin (1:1000) and actin clone C4 (1:5000), with or without rabbit polyclonal Lamin A/C antibody (1:3000, sc-20681; Santa Cruz Biotechnology). Relative intensity was calculated using the ratio of each target protein's signal intensity to internal controls' intensity, using ImageJ ver1.52p (NIH, Bethesda, MD, USA). At least three separate experiments were conducted. Full-length blots were absent because membranes were cut prior to hybridization with primary antibodies. The unedited blots including replicates were shown in Supplementary Figs.S5 , S6 , S7 .
Western blotting was performed as previously described 45 (link) with the following primary antibodies: β-catenin (1:1000) and actin clone C4 (1:5000), with or without rabbit polyclonal Lamin A/C antibody (1:3000, sc-20681; Santa Cruz Biotechnology). Relative intensity was calculated using the ratio of each target protein's signal intensity to internal controls' intensity, using ImageJ ver1.52p (NIH, Bethesda, MD, USA). At least three separate experiments were conducted. Full-length blots were absent because membranes were cut prior to hybridization with primary antibodies. The unedited blots including replicates were shown in Supplementary Figs.
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