Fig 2: The role of M-Sec and RalA in TNT formation in OCI-AML3 cellsA-B. OCI-AML3 cells transduced with shCtr, sh-M-Sec and shRalA. Representative immunoblots of three independent experiments are shown for verification of shRNA knock-down of M-Sec (A) and RalA (B). Immunoblots were quantified with Image Quant TL with significant changes indicated (**P < 0.01). C. TNT quantifications were performed in parallel with the three independent immunoblot experiments in A-B. Data are displayed as mean ± S.D. n.s. = not significant

Fig 3: Investigation of the molecular machinery in TNT development in AML cell linesA. AML cells were lysed and subjected to immunoblotting using antibodies against M-Sec, RalA, LST1 and GAPDH (loading control). Representative blots from three independent experiments are shown. B. OCI-AML3 cells were fixed and immunostained with antibodies against M-Sec, LST1 and RalA and analyzed by fluorescence microscopy. C. Arrow indicates localization of M-Sec in TNT-like structures. D. NB4, MOLM-13 and HL60 cells were treated for 24 h with 0.15% EtOH (Ctr), ATRA (10 µM) or TNFa (10 nM) alone or in combination and immunoblotted using antibodies against M-Sec, RalA, LST1 and GAPDH as loading control. Representative blots from three independent experiments are shown. E. Quantification of TNTs in NB4, MOLM-13 and HL60 cells (n = 3) (*P < 0.05, **P < 0.01). All scale bars = 10 µm.

Fig 4: The NF-?B pathway is involved in TNT formation in AML cellsA. AML cell lines were lysed and immunoblotted using antibodies against NF-?B-p65 and COXIV as loading control. Representative blots from three independent experiments are shown. B. OCI-AML3 cells were treated with BAY 11-7082 (2.5 µM, 24 h) and analyzed by immunoblotting with antibodies against NF-?B p65 and NF-?B p-p65. Representative blots from three independent experiments are shown. C. TNT quantification of OCI-AML3 cells treated with BAY 11-7082 (2.5 µM, 24 h) and control (n = 3) (**P < 0.01). D. OCI-AML3 cells were treated with BAY 11-7082 (2.5 µM, 24 h) and analyzed by immunofluorescence with antibodies against RalA, M-Sec and LST1. E. OCI-AML3 cells were treated with BAY 11-7082 (2.5 µM, 24 h) and immunoblotted with antibodies against RalA, M-Sec and LST1 with additional GAPDH as a loading control. Representative blots from three independent experiments are shown in addition to the quantification. Scale bars = 10 µm. Data are displayed as mean ± S.D.

Fig 5: p-STAT1 may recruit EP300 to promote target gene expression in NCM460 cells. a EP300 mRNA expression in DSS-induced mice chronic colitis (error bars: mean ± SEM; control group n = 7, DSS group n = 9). b EP300 mRNA expression in IBD patients (error bars: mean ± SEM; NC group n = 8, UC group n = 8, CD group n = 10). c H3K27ac protein expression in NCM460 after adding EP300 inhibitor C646 in different concentration (0, 10, 20, 30, 40, 50 µM) for 24 h. d–e LCP2 and TNFAIP2 mRNA expression in NCM460 after adding C646 or DMSO without or with stimulation of by TNF-a (error bars: mean ± SD). f–g EP300 mRNA and protein expression in NCM460 after transfected with siEP300-1, 2, 3 or the negative control (error bars: mean ± SD). h–i LCP2 and TNFAIP2 mRNA expression in NCM460 after transfected with siEP300 or the negative control with or without stimulation by TNF-a (error bars: mean ± SD). j LCP2 and TNFAIP2 mRNA expression in NCM460 after transfected with siEP300 or the negative control with stimulation by TNF-a + IFN-? (error bars: mean ± SD). k LCP2 and TNFAIP2 mRNA expression in NCM460 after transfected with siSTAT1 + siEP300 or the negative control with stimulation by TNF-a + IFN-? (error bars: mean ± SD). l co-IP experiment was performed to validate the combination between p-STAT1 and EP300. *p < 0.05, **p < 0.01, ***p < 0.001, ns means no significance