Fig. 1
HNRNPA2B1 promotes MM proliferation and increases ILF3 protein expression through recognizing m6A modification and maintaining stabilization of ILF3 mRNA. a HNRNPA2B1 mRNA levels were significantly increased in MM samples. The signal level of HNRNPA2B1 was shown on the y-axis. Patients were designated as healthy donors with normal bone marrow plasma cells (NP, n = 22), monoclonal gammopathy of undetermined significance (MGUS, n = 44), or multiple myeloma (MM, n = 351), which were sorted on the x-axis. b Elevated HNRNPA2B1 mRNA was associated with poor overall survival (OS) in MM patients from the APEX patient cohort. c Overexpression of HNRNPA2B1 was confirmed by Western blotting after lentivirus infection in MM cells. d Overexpression of HNRNPA2B1 promoted cell proliferation in ARP1 and H929 cells. e Flow cytometry showed that HNRNPA2B1 inhibited MM cells apoptosis. f Expression of HNRNPA2B1 and apoptosis-related proteins in HNRNPA2B1KD MM cells. g Tumor tissues were dissected from NOD-SCID mice injected with wild-type and HNRNPA2B1OE cells. h Hot map of 44 differentially expressed genes (P < 0.05) in both m6A and transcription in ARP1 HNRNPA2B1KD cells compared with controls. i HORMER motif analysis of m6A peaks in ARP1 and H929 cells. "Target" showed the percentage of peaks containing the identified consensus motif, "Background" presented the percentage of genome background regions that contain the identified motif. j IGV tracks depicted the position of m6A modification of ILF3 gene. k HNRNPA2B1 gene knockdown affected expression of ILF3 mRNA in MM cells. l RIP-qPCR assay was used to determine the interaction between HNRNPA2B1 and ILF3 mRNA in ARP1 cells. m The expression of ILF3 RNA was detected in ARP1 cells upon treating with cyclolencine in the concentration of 0 mM, 50 mM and 100 mM. n RT-qPCR following the addition of Actinomycin D (5 μg/mL) was performed to detect ILF3 mRNA stability. k HNRNPA2B1 (red) localization was examined by confocal microscopy. (i) Nuclei were stained with DAPI. Scale bar, 5 μm. (ii) Immunolocalization of HNRNPA2B1 in MM cells. (iii) Immunolocalization of ILF3. (iv) Merger of images of i, ii and iii, with the colocalized regions shown in orange. (*P < 0.05, **P < 0.01, ***P < 0.001)