Fig. 2

BCL7A restoration promotes a tumor suppressor-like phenotype. a Differential expression of BCL7A mRNA between acute myeloid leukemia (AML) and diffuse large B-cell lymphoma (DLBCL) cell lines from DepMap. TPM: transcripts per million (Supplementary Table 1). b Methylation Specific PCR (MSP). PCR products generated by the specific primers for the methylated sequence (M) and for the unmethylated sequence (U) are depicted. MSP was carried out using genomic DNA from the NB4 cell line, as well as completely methylated and unmethylated genomic DNAs. In addition, a negative control with no DNA template was included. c Relative BCL7A protein expression differences of the NB4 cell line due to Decitabine (DAC) treatment. BCL7A presents two alternative isoforms that are co-expressed due to an extended exon variant located in exon 5, and due to this, there are two bands for BCL7A. β-actin was used as a loading control and the numeric result was normalized with this loading control signal. d Competition cell growth effect of BCL7A expression restoration on in vitro proliferation. Competition cell growth assay of non-transduced NB4 cells vs NB4 cells transduced with either pLVX-IRES-ZsGreen1 plasmid expressing wild-type BCL7A, mutant BCL7A (∆27-BCL7A), or empty vector. All constructs were evaluated in a time course of ZsGreen1.⁺ % measurements on days 1, 5, 12, 19, and 26. This assay involves the co-culture of an NB4 model transduced with one of the plasmids together with non-transduced NB4 cells. e Bioluminescence signal at day 21 after injection of NB4-luciferase-transduced cells in the empty vector mice group (n = 6) and wild-type BCL7A mice group (n = 6) groups. f Image of the bioluminescence signal of two representative NGS mice in ventral position on day 21 after the tail-injection of NB4-luciferase cells transduced with BCL7A wildtype (up) and empty vector (down)