Fig 2: Activation of CD19 and CD79b expression in KIS-1 following doxycycline-induced expression of EBF1 for 24–96 h. Activation of CD19 (a) and CD79b (b) shown by RT-qPCR. Activation of CD79b expression shown by Western blotting (c). Data represents 3 independent experiments.

Fig 3: Confirmation of PAX5 interacting proteins in KIS-1 and REH. (a) Western Blot to show EBF1 and PAX5 expression in KIS-1 + EBF1 nuclear and cytoplasmic (cyto) extracts with and without DOX-induction for 48 h. (b) Coomassie Blue-staining of PAX5 co-immunoprecipitated material from KIS-1 + EBF1 nuclear extracts with or without doxycycline induction for 48 h. (c) Western blot to show KMT2A co-immunoprecipitation with PAX5 from KIS-1 + EBF1 nuclear extracts with and without doxycycline induction for 48 h. (d) Western blot to show MEN1 co-immunoprecipitation with PAX5 from REH and GM12878 nuclear extracts.

Fig 4: Ebf1 overexpression attenuates LPS-induced inflammation, oxidative stress, impaired proliferation, and apoptosis in MLE-15 cells. MLE-15 cells were transduced with control lentivirus or Ebf1-overexpressing lentivirus. a, b The expression levels of Ebf1 mRNA (a) and protein (b) were quantitated by RT-qPCR and western blot assays, respectively. c–g MLE-15 cells after transfection with control lentivirus or Ebf1-expressing virus were further cultured in medium supplemented with PBS or 100 ng/mL LPS. c The TNF-α and IL-6 levels in MLE-15 cell culture supernatant was quantitated by ELISA. d Cellular ROS was determined by flow cytometric analysis of DCFDA staining. e Cell proliferation was determined by CCK-8 assay. f, g Apoptosis was determined by TUNEL staining. Scale bar, 100 µm. * p < 0.05, ** p < 0.01, *** p < 0.001

Fig 5: Ebf1 is a downstream target of mmu-miR-598, and Ebf1 expression in MLE-15 cells is downregulated by LPS stimulation. a Sequence alignment of Ebf1 3'UTR and miR-598. WT, wild type; MUT, mutated. b Immortalized mouse lung epithelial MLE-15 cells were transfected with luciferase reporter vector containing WT or mutated 3′-UTR of Ebf1, together with miR-598 mimics or negative control. The dual luciferase assay was performed to validate the interaction between miR-598 and Ebf1. c, d MLE-15 cells were transfected with miR-598 mimics, miR-598 specific inhibitor, or relative negative control miRNA. At 48 h after transfection, cells were harvested to analyze the mRNA and protein levels of Ebf1 by qPCR (c) and western blot (WB) (d), respectively. e, f MLE-15 cells were treated with different concentration of LPS (e) or treated with LPS for different durations, and the relative expression of Ebf1 mRNA was quantitated by RT-qPCR. g The relative expression level of Ebf1 in lung tissues of sham/LPS treated mice at 24 h after challenge was determined by RT-qPCR. * p < 0.05, ** p < 0.01, *** p < 0.001