Journal: Cell Communication and Signaling : CCS

Article Title: Cytoplasmic retention of IRF3 binding Vimentin competitively with ERK1/2 mitigates acute myeloid leukemia through TFEB nuclear translocation

doi: 10.1186/s12964-026-02899-9

Figure Lengend Snippet: IRF3-Vimentin interaction impedes Vimentin-p-ERK signaling activation leading to PPP3CB-mediated dephosphorylation of TFEB and its nuclear translocation. A - C Co-immunoprecipitation of Vimentin, ERK and p-ERK was performed to confirm their interaction in THP-1 cells with PMA stimulation for 24 h. D The molecular docking simulations between Vimentin and ERK by AlphaFold3. Structural visualization was done using PyMOL. E Co-immunoprecipitation between Flag-ERK2 or Flag-ERK1 and V5-IRF3 or HA-Vimentin were evaluated after three plasmids co-transfection in 293 T cells for 48 h. F Co-immunoprecipitation between HA-Vimentin or Flag-ERK2 and V5-IRF3 after three plasmids co-transfection in 293 T cell. G p-ERK and ERK expression in IRF3 knockdown cell line with PMA stimulation 24 h was detected by Western blotting. GAPDH was used as housekeeping control. Experiments were performed at least three independent times, and representative results are shown. H Co-immunoprecipitation of Vimentin and p-ERK or ERK in IRF3 knockdown cell line was performed to confirm their interaction in THP-1 cells with PMA stimulation 24 h. I p-ERK, ERK, TFEB, and PPP3CB expression in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h was detected by Western blotting. GAPDH was used as housekeeping control. Experiments were performed at least three independent times, and representative results are shown. J Co-immunoprecipitation of Vimentin and p-ERK or ERK in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h. K Co-immunoprecipitation of ERK and TFEB in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h. L Co-immunoprecipitation of PPP3CB and TFEB in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h. M The expression of TFEB in cytoplasm and nucleus in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h was detected by Western blotting. GAPDH was used as housekeeping control in cytoplasm and Lamin B1 in nucleus. The rightmost control sample was nuclear or cytoplasm protein which was used as the control. Experiments were performed at least three independent times, and representative results are shown. M – O The IRF3 overexpressing cell line was stimulated with 1 μM TA1 ( n = 6), 10 μM PD98059 ( n = 6), or DMSO control ( n = 6) for 48 h, and the expression levels of CD14 and CD11b were examined via flow cytometry. Experiments were performed at least three independent times, and representative results are shown. The statistical results and representative plots are shown. * P < 0.05, ** P < 0.01 by one-way ANOVA and post hoc Bonferroni test

Article Snippet: The IRF3 inhibitor Amlexanox (25 μM, MedChemExpress, HY-B0713), TFEB activator TA1 (1 μM), or ERK1/2 inhibitor PD98059 (10 μM), were administered to THP-1 cells, or IRF3-overexpressed THP-1 cells for 48 h in the presence or absence of PMA stimulation.

Techniques: Activation Assay, De-Phosphorylation Assay, Translocation Assay, Immunoprecipitation, Cotransfection, Expressing, Knockdown, Western Blot, Control, Mutagenesis, Flow Cytometry

Journal: Cell Communication and Signaling : CCS

Article Title: Cytoplasmic retention of IRF3 binding Vimentin competitively with ERK1/2 mitigates acute myeloid leukemia through TFEB nuclear translocation

doi: 10.1186/s12964-026-02899-9

Figure Lengend Snippet: TA1 and PD98059 decrease leukemia burden in a disseminated intravenous xenograft leukemia model induced by IRF3 up-regulation. A A scheme of designing TA1 and PD98059 treatment process in pLJM1-IRF3 cell induced disseminated intravenous xenograft leukemia model. B - C The leukemia burden among Control ( n = 9), TA1 ( n = 9) and PD98059 ( n = 10) group in the peripheral blood ( B ) and bone marrow ( C ) of NOD/SCID mice are shown as statistical analysis and representative flow cytometry plot. Data are shown as mean ± SEM and *** P < 0.001 by Mann–Whitney U test. D The clinical leukemia scores among Control ( n = 10), TA1 ( n = 10) and PD98059 ( n = 10) group were graded on days 30 after the transplantation. Data are shown as mean ± SEM and *** P < 0.001 by Mann–Whitney U test. E – F The leukemia cells of human CD45 + in the peripheral blood ( E ) and bone marrow ( F ) among Control ( n = 10), TA1 ( n = 10) and PD98059 ( n = 10) group were stained by Wright-Giemsa staining and the leukemia burden was shown as statistical analysis. Data are shown as mean ± SEM and *** P < 0.001 by Mann–Whitney U test (Scale bars: 200 μm). G Proposed model showing how IRF3 cytoplasmic retention regulates AML cell differentiation. PAMPs activate IRF3, promoting inflammation and proliferation. With pro-differentiation agent (PDA, e.g., PMA), IRF3 is retained in the cytoplasm, interacts with Vimentin, disrupts Vimentin-p-ERK signaling, and enhances TFEB-mediated differentiation gene expression

Article Snippet: The IRF3 inhibitor Amlexanox (25 μM, MedChemExpress, HY-B0713), TFEB activator TA1 (1 μM), or ERK1/2 inhibitor PD98059 (10 μM), were administered to THP-1 cells, or IRF3-overexpressed THP-1 cells for 48 h in the presence or absence of PMA stimulation.

Techniques: Control, Flow Cytometry, MANN-WHITNEY, Transplantation Assay, Staining, Cell Differentiation, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Cytoplasmic retention of IRF3 binding Vimentin competitively with ERK1/2 mitigates acute myeloid leukemia through TFEB nuclear translocation

doi: 10.1186/s12964-026-02899-9

Figure Lengend Snippet: IRF3-Vimentin interaction impedes Vimentin-p-ERK signaling activation leading to PPP3CB-mediated dephosphorylation of TFEB and its nuclear translocation. A - C Co-immunoprecipitation of Vimentin, ERK and p-ERK was performed to confirm their interaction in THP-1 cells with PMA stimulation for 24 h. D The molecular docking simulations between Vimentin and ERK by AlphaFold3. Structural visualization was done using PyMOL. E Co-immunoprecipitation between Flag-ERK2 or Flag-ERK1 and V5-IRF3 or HA-Vimentin were evaluated after three plasmids co-transfection in 293 T cells for 48 h. F Co-immunoprecipitation between HA-Vimentin or Flag-ERK2 and V5-IRF3 after three plasmids co-transfection in 293 T cell. G p-ERK and ERK expression in IRF3 knockdown cell line with PMA stimulation 24 h was detected by Western blotting. GAPDH was used as housekeeping control. Experiments were performed at least three independent times, and representative results are shown. H Co-immunoprecipitation of Vimentin and p-ERK or ERK in IRF3 knockdown cell line was performed to confirm their interaction in THP-1 cells with PMA stimulation 24 h. I p-ERK, ERK, TFEB, and PPP3CB expression in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h was detected by Western blotting. GAPDH was used as housekeeping control. Experiments were performed at least three independent times, and representative results are shown. J Co-immunoprecipitation of Vimentin and p-ERK or ERK in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h. K Co-immunoprecipitation of ERK and TFEB in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h. L Co-immunoprecipitation of PPP3CB and TFEB in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h. M The expression of TFEB in cytoplasm and nucleus in IRF3 S386A and S386E mutant cell line with PMA stimulation 24 h was detected by Western blotting. GAPDH was used as housekeeping control in cytoplasm and Lamin B1 in nucleus. The rightmost control sample was nuclear or cytoplasm protein which was used as the control. Experiments were performed at least three independent times, and representative results are shown. M – O The IRF3 overexpressing cell line was stimulated with 1 μM TA1 ( n = 6), 10 μM PD98059 ( n = 6), or DMSO control ( n = 6) for 48 h, and the expression levels of CD14 and CD11b were examined via flow cytometry. Experiments were performed at least three independent times, and representative results are shown. The statistical results and representative plots are shown. * P < 0.05, ** P < 0.01 by one-way ANOVA and post hoc Bonferroni test

Article Snippet: In experiments involving treatment with the ERK1/2 inhibitor (PD98059, MedChemExpress, HY-12028) and TFEB activator (TA1, MedChemExpress, HY-135825), mice were randomly assigned to different treatment groups at 14 days post leukemia cell transplantation.

Techniques: Activation Assay, De-Phosphorylation Assay, Translocation Assay, Immunoprecipitation, Cotransfection, Expressing, Knockdown, Western Blot, Control, Mutagenesis, Flow Cytometry

Journal: Cell Communication and Signaling : CCS

Article Title: Cytoplasmic retention of IRF3 binding Vimentin competitively with ERK1/2 mitigates acute myeloid leukemia through TFEB nuclear translocation

doi: 10.1186/s12964-026-02899-9

Figure Lengend Snippet: TA1 and PD98059 decrease leukemia burden in a disseminated intravenous xenograft leukemia model induced by IRF3 up-regulation. A A scheme of designing TA1 and PD98059 treatment process in pLJM1-IRF3 cell induced disseminated intravenous xenograft leukemia model. B - C The leukemia burden among Control ( n = 9), TA1 ( n = 9) and PD98059 ( n = 10) group in the peripheral blood ( B ) and bone marrow ( C ) of NOD/SCID mice are shown as statistical analysis and representative flow cytometry plot. Data are shown as mean ± SEM and *** P < 0.001 by Mann–Whitney U test. D The clinical leukemia scores among Control ( n = 10), TA1 ( n = 10) and PD98059 ( n = 10) group were graded on days 30 after the transplantation. Data are shown as mean ± SEM and *** P < 0.001 by Mann–Whitney U test. E – F The leukemia cells of human CD45 + in the peripheral blood ( E ) and bone marrow ( F ) among Control ( n = 10), TA1 ( n = 10) and PD98059 ( n = 10) group were stained by Wright-Giemsa staining and the leukemia burden was shown as statistical analysis. Data are shown as mean ± SEM and *** P < 0.001 by Mann–Whitney U test (Scale bars: 200 μm). G Proposed model showing how IRF3 cytoplasmic retention regulates AML cell differentiation. PAMPs activate IRF3, promoting inflammation and proliferation. With pro-differentiation agent (PDA, e.g., PMA), IRF3 is retained in the cytoplasm, interacts with Vimentin, disrupts Vimentin-p-ERK signaling, and enhances TFEB-mediated differentiation gene expression

Article Snippet: In experiments involving treatment with the ERK1/2 inhibitor (PD98059, MedChemExpress, HY-12028) and TFEB activator (TA1, MedChemExpress, HY-135825), mice were randomly assigned to different treatment groups at 14 days post leukemia cell transplantation.

Techniques: Control, Flow Cytometry, MANN-WHITNEY, Transplantation Assay, Staining, Cell Differentiation, Gene Expression