Extended Data Fig. 4: RBMX and RBMXL1 are required for human myeloid leukemia cell survival.
a, Giemsa staining of control and RBMX/L1-knockdown MOLM13 cells from Fig. 4k. Original magnification 400X. Scale bar, 20μm. The experiment was performed once; control and shRNA1: 9 images, shRNA2: 16 images were collected. (b-d) Representative FACS plots of Fig. 4m, o and p, respectively. e, Immunoblot analysis of RBMX/L1 in KCL-22 transduced with EV or RBMX overexpressing vector (RBMX-R2) and shRNA control or shRNAs against RBMX/L1. The experiment was performed 3 times with similar results. f, Proliferation assay of cells in e. n = 3 independent experiments. g, Quantitative FACS analysis of apoptotic cells in control and RBMX/L1-knockdown KCL-22-EV and KCL-22-RBMX-R2 cells 5 days post transduction. n = 3 independent experiments. h, Immunoblots for RBMX/L1 in MOLM13-Cas9 cells transduced with sgRNAs targeting RBMX/L1 (sg-1 and sg-2). The experiment was performed 4 times with similar results. i, Proliferation assay of cells in h. n = 4 independent experiments. j, FACS analysis of myeloid differentiation markers CD11b and CD33 in cells from h. n = 4 independent experiments. k, Quantitative FACS analysis summary of apoptosis of cells in h, 3 days post transduction. n = 4 independent experiments. l, Immunoblots analysis of GFP+ PDX AML-1 cells. The experiment was performed once. m, qRT-PCR showing depletion of RBMX/L1 in GFP+ PDX AML-13 cells. n = 1 experiment. n, Immunoblots for RBMX/L1 upon RBMX/L1 depletion in GFP+ PDX AML-11 cells. The experiment was performed once. o, Representative FACS plots of Fig. 4t. p, Immunoblot and band densitometry of RBMX/L1 in BM cells from animals transplanted with PDX AML-1 cells that succumbed to leukemia. The experiment was performed once. (q-s) Representative FACS plots of Fig. 5c, g and h, respectively. Data presented in f-g, i-k as mean, error bars, s.e.m. p values were calculated using two-tailed Student’s t test, unless indicated otherwise.
