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. 2021 Aug 31;36(2):438–451. doi: 10.1038/s41375-021-01394-z

Fig. 2. Knockdown of USP15 impairs human leukemic cell function in vitro and in vivo.

Fig. 2

A Relative proportion of the indicated AML cell lines transduced with shRNAs targeting USP15 (shUSP15) or a scrambled control shRNA (shSCR) co-expressing mCherry. The percent of mCherry+ cells was initially determined by flow cytometry at 48 h after transduction (Day 0). The proportion of mCherry+ cells was normalized to the percent mCherry+ on Day 0 for each group. Data reflects two independent assays (n = 3 per group). B Total cell number of OCI-AML3 cells transduced with shRNAs targeting USP15 (shUSP15) or a scrambled control shRNA (shSCR). *P < 0.05. Data reflects two independent assays (n = 3 per group). C The relative number of colonies formed in methylcellulose from the indicated AML cells transduced with shRNA targeting USP15 (shUSP15) or a scrambled control shRNA (shSCR) co-expressing mCherry. Cells were sorted for mCherry expression prior to plating. *P < 0.05; **P < 0.005; ns, not significant. (n = 3) Data reflects two independent assays. D The total number of colonies formed in methylcellulose from healthy CD34+ cells transduced with shRNAs targeting USP15 (shUSP15) or a scrambled control shRNA (shSCR) co-expressing mCherry (n = 3 per group). *P < 0.05. E Schematic depicting unsorted OCI-AML3 cells transduced with shRNAs targeting USP15 (shUSP15) or a scrambled control shRNA (shSCR) xenografted into NSGS mice. F Percent of human CD45+populations expressing shSCR (n = 7 mice per group) or shUSP15 (n = 8 mice per group) (mCherry+) was evaluated in the bone marrow, peripheral blood, and spleen at the time of death. *P < 0.05.