Fig. 6: IL-17A activates NF-kB to induce CXCL16 transcription and CXCL16 secretion in leukemia cells. | Nature Communications

Fig. 6: IL-17A activates NF-kB to induce CXCL16 transcription and CXCL16 secretion in leukemia cells.

From: Targeting IL-17A enhances imatinib efficacy in Philadelphia chromosome-positive B-cell acute lymphoblastic leukemia

Fig. 6: IL-17A activates NF-kB to induce CXCL16 transcription and CXCL16 secretion in leukemia cells.The alternative text for this image may have been generated using AI.

Primary Ph+ B-ALL cells were treated with or without rhIL-17A for 24 h, and relative CXCL16 mRNA level (a) and CXCL16 secretion (b) were evaluated by real-time PCR and ELISA, respectively. c Representative immunofluorescence images of B220 (red) and CXCL16 (green) staining in spleen tissue sections of WT mice and BCR-ABLtTA mice treated with or without 50 µg/kg rmIL-17A for a duration of 3 weeks (twice a week) (n = 3 mice per group). d Flow cytometric analysis and quantification of CXCL16 expression in primary mouse B-ALL cells treated with or without rmIL-17A. e The protein levels of p65 and p-p65 in the cytoplasm and p-p65 in the nucleus of primary mouse leukemia cells treated with or without rmIL-17A were measured by Western blotting. f Immunofluorescence of p-P65 in primary B-ALL cells treated with or without rhIL-17A. g The effect of rhIL-17A treatment on NF-kB transcriptional activity. HEK 293T cells were transfected with a synthetic NF-kB luciferase reporter construct (pNF-kB-Luc) for 12 h and then treated with different concentrations of rhIL-17A for 24 h. NF-kB transcriptional activity was detected by a luciferase assay. h ChIP‒qPCR analyses of the binding of NF-kB to the CXCL16 promoter region in SupB15 cells treated with or without rhIL-17A. i–k The effect of BAY11-7082, rIL-17A or BAY11-7082 in combination with rIL-17A on the cytoplasmic and nuclear protein levels of p65, p-p65, and CXCL16 in primary Ph+ B-ALL cells. i The indicated protein band intensities were quantified using ImageJ software. The relative CXCL16 mRNA level (j) in primary Ph+ B-ALL cells and CXCL16 protein level (k) in the supernatant of primary Ph+ B-ALL cells were measured by RT‒PCR and ELISA, respectively. (a, b, d–k) n = 3 independent experiments. Statistical significance was calculated by (a, b, d) two-tailed Student’s t-test; (e, g–k) one-way ANOVA with Tukey’s multiple comparison tests; Data are presented as means ± S.E.M. Source data are provided as a Source Data file.

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