Prior to mass spectrometry analysis, samples were desalted using a 96-well plate filter (Orochem) packed with 1 mg of Oasis HLB C-18 resin (Waters). Briefly, the samples were resuspended in 100 µl of 0.1% TFA and loaded onto the HLB resin, which was previously equilibrated using 100 ...
To evaluate the role of KDM2B protein domains we exploited the fact that in-frame deletions produced by CRISPR-Cas9-directed mutagenesis (which occur in ∼1/3 cases) are more deleterious to protein function when they disrupt critical domains (Shi et al., 2015); consequently, in competition ...
Upon shRNA-induced knockdown of KDM1A, a moderate but significant decrease in cell viability was detected in all tested primary tumor-derived CRC-SCs (n = 4) and HCT116 cells, with more than 20% inhibition of cell viability over a timeframe of 9 days (Fig. 2B, C). Fig. 2: ...