Fig. 1: Identification of UBE2J2 and UBE2K as the collaborating E2 proteins of MARCH5. | Leukemia

Fig. 1: Identification of UBE2J2 and UBE2K as the collaborating E2 proteins of MARCH5.

From: The UBE2J2/UBE2K-MARCH5 ubiquitination machinery regulates apoptosis in response to venetoclax in acute myeloid leukemia

Fig. 1

a Scatterplot showing the log2 fold-change (LFC) of sgRNA abundance and the associated p-value in venetoclax-treated cells compared to DMSO control cells. The scores of all sgRNAs targeting the same gene were averaged and shown as one data point. b Relative viability of AML cells expressing the indicated sgRNAs after 3-day venetoclax (VEN) treatment. Cell viability was determined by CellTiter-Glo and normalized to the DMSO-treated control. The mean ± SD (n = 4) and dose-response curves are plotted. sgNT, a non-targeting sgRNA. c Scatterplot showing the Pearson correlations between the dependency scores of MARCH5 and UBE2J2 or UBE2K across AML cell lines or other cancer cell lines in the DepMap dataset. Each dot represents a cell line. d In vitro competitive growth of AML cells co-expressing the indicated sgRNAs and GFP. Relative cell growth is measured by the change in the percentage of GFP+ cells; results represent mean + SD, n = 2. e Competitive growth of PDX17-14 cultured cells transduced with an empty vector (Ctrl) or the CRISPR-resistant cDNA of UBE2J2 or UBE2K upon the knockout of the corresponding endogenous E2 genes; results represent mean + SD, n = 2. f Immunoblot analysis of MCL1 and NOXA upon MARCH5, UBE2J2 or UBE2K knockout in MOLM14 cells. g Immunoblot analysis confirming the expression of LgBiT-tagged MARCH5 proteins and SmBiT-tagged E2 proteins in transduced MV4-11 cells. Non-transduced cells were used as a negative control (neg ctrl). h Relative luminescent signals of MV4-11 cells expressing the indicated NanoBiT constructs; results represent mean ± SD, n = 4. ***p < 0.001; ****p < 0.0001, unpaired two-sided t-test. i. Relative cell viability with venetoclax treatment (mean ± SD, n = 3) of MV4-11 cells transduced with the indicated sgRNA combinations. Immunoblot analysis to confirm the gene knockout j and competition proliferation assays to evaluate the growth effect of E2 depletion as single or double-knockout in MV4-11 (mean + SD, n = 2) k.

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