recombinant murine il 4  (R&D Systems)

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 impairs early B-cell development in the bone marrow. (A) Experimental scheme for AAV-mediated IL-27 delivery and analysis of splenic and BM B-cell compartments. (B–D) Splenic B-cell analysis 10 weeks after AAV-Ctrl or AAV-IL-27 treatment (n = 2 per group). (B) Frequency of total splenic B cells. (C) CIBERSORTx-based deconvolution of splenic B-cell subsets, including clustering and pseudotime analysis (left) and relative subset distribution (right). (D) Cd93 expression (TPM) in splenic B cells. (E–G) Analysis of splenic and BM B-cell compartments 3 weeks after treatment (n = 4 per group). (E) Representative flow cytometry plots. (F) Frequencies of B220 + CD19 − and B220 − CD19 + populations in the BM and spleen. (G) Relative distribution of B-cell developmental subsets in the BM. (H–I) Kinetics of B-cell populations following IL-27 treatment (n = 2 per group at each time point). (H) Frequency and absolute number of BM B220 + B cells. (I) Proportions of B-cell subsets within the B220 + compartment at the indicated time points. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Cell Analysis, Expressing, Flow Cytometry

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 inhibits CLP formation in the bone marrow. (A) qPCR analysis of B-lineage- and myeloid-associated transcription factors in BM cells 5 days after AAV-Ctrl or AAV-IL-27 administration (n = 3 per group). (B) qPCR analysis of CEBPA expression in Reh cells treated with hIL-27 and DAC for 3 days (n = 3 per group). (C–D) Frequencies and absolute numbers of CLPs in WT and CD19 Cre ;IL-27R fl/fl mice (n = 3 per group). (E–F) BM B-cell frequency (E) and proportion of B220 + CD43 hi IgM − B cells (F) in WT and CD19 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for WT Ctrl group; n = 3 for WT IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 4 for CD19 Cre IL-27 group). (G–H) BM B-cell frequency (G) and proportion of B220 + CD43 hi IgM − B cells (H) in CD4 Cre ;IL-27R fl/fl and Lyz2 Cre ;IL-27R fl/fl mice 2 weeks after AAV treatment (n = 3 for CD4 Cre Ctrl group; n = 3 for CD4 Cre IL-27 group; n = 5 for Lyz2 Cre Ctrl group; n = 6 for Lyz2 Cre IL-27 group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 reshapes immune reconstitution after bone marrow transplantation. (A) Experimental scheme. WT mice underwent BM transplantation followed by AAV-Ctrl or AAV-IL-27 treatment. Splenic and BM immune reconstitution was analysed at 10 weeks after treatment (n = 4 mice for the Ctrl group; n = 5 mice for the IL-27 group). (B) Percentages and absolute numbers of CD45 + splenocytes. (C–F) Frequencies of major immune cell populations within CD45 + splenocytes: myeloid cells (C), macrophages (D), T and B cells (E), and NK cells (F). (G) Frequency of CD4 + T cells among splenic T cells. (H) Proportion of Tregs among CD4 + T cells. (I–J) BM B-cell reconstitution. Frequency of BM B220 + B cells (I) and distribution of B220 + B-cell subsets (J). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 inhibits B-cell development through both direct and indirect mechanisms. (A) Schematic of the BM chimera model. WT (WTR) or IL-27R −/− (KOR) recipients were treated with AAV-Ctrl or AAV-IL-27. Donor BM cells were a 1:1 mix of WT (CD45.1 + ) and IL-27R −/− (CD45.1 − ) cells. Analyses were performed at 5 weeks (B–E; n = 3 per group) or 10 weeks (F–I; n = 5 per group) post-transplantation. (B) Percentage of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (C) Absolute numbers of total CD19 + B cells in spleen and BM of WTR and KOR mice. (D) Absolute numbers of CD19 + B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in spleen and BM of WTR and KOR mice. (E) Percentage of B220 + CD19 − B cells from CD45.1 + or CD45.1 − (IL-27R −/− ) donors in BM of WTR and KOR mice. (F) Absolute numbers of B220 + CD19 - B cells from CD45.1 + or CD45.1 - (IL-27R −/− ) donors in BM of WTR and KOR mice. (G) Frequencies and absolute numbers of BM B220 + B cells derived from CD45.1 + or IL-27R −/− donors in WTR mice. (H) Proportion of CD43 hi IgM − subsets from CD45.1 + or IL-27R −/− donors in BM B220 + B cells in WTR mice. (I–J) Frequencies of CLPs (I) and their donor-specific contributions (J) in BM chimeras. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Transplantation Assay, Derivative Assay

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: Exogenous IL-27 indirectly inhibits BM B-ALL cells. (A) Experimental design. GFP + B-ALL cells were transferred into WT mice, followed by AAV-Ctrl or AAV-IL-27 administration. BM and splenic B-ALL cells were analysed 9–10 days later (n = 3 per group). (B–C) Representative flow cytometry plots and quantification of GFP + B-ALL and GFP − B cells. (D) GSVA analysis of transcriptional changes in BM B-ALL cells isolated from mice in (A). (E) Gene Ontology (GO) enrichment analysis of DEGs between Ctrl and IL-27 groups in (A) (adjusted P < 0.05, |log 2 fold change| > 1). The outer ring indicates enriched GO terms, the middle ring represents relative gene expression changes in the IL-27 group, and the inner ring shows background gene counts, with colour intensity reflecting enrichment significance. Enriched pathways include interferon-γ-mediated signalling, interferon-β responses, adhesion-related processes, protozoan defence responses, calmodulin-dependent kinase signalling, and negative regulation of STAT tyrosine phosphorylation. (F–G) Expression of apoptosis- and cell cycle-related genes in BM B-ALL cells from (A). (H) Experimental design. WT and IL-27R −/− mice transplanted with GFP + B-ALL cells were treated with AAV-Ctrl or AAV-IL-27 (n = 4 for WT Ctrl group; n = 4 for WT IL-27 group; n = 5 for IL-27R −/− Ctrl group; n = 5 for IL-27R −/− IL-27 group). (I–J) Representative flow plots and quantification of BM B-ALL cells. (K–L) Flow cytometry plots and quantification of BM B-ALL cells in CD4 Cre ;IL-27R fl/fl , CD19 Cre ;IL-27R fl/fl , and Lyz2 Cre ;IL-27R fl/fl mice (n = 2 for CD4 Cre Ctrl group; n = 2 for CD4 Cre IL-27 group; n = 3 for CD19 Cre Ctrl group; n = 3 for CD19 Cre IL-27 group; n = 3 for Lyz2 Cre Ctrl group; n = 3 for Lyz2 Cre IL-27 group). (M) Flow cytometry plots and quantification of BM B-ALL cells in Rag1 −/− mice (n = 3 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Flow Cytometry, Isolation, Gene Expression, Phospho-proteomics, Expressing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 reshapes the BM niche, reducing B-cell support and leukaemogenesis. (A) Pseudotime analysis of BM B-cell subsets (C0–C10). (B) Expression of representative marker genes across B-cell clusters in (A). (C–G) BM cells were harvested 8 days after AAV-Ctrl or AAV-IL-27 administration for RNA-seq analysis (n = 3 per group). (C) CIBERSORTx analysis showing increased pre-pro-B and cycling pro-B subsets after IL-27 treatment. (D) KEGG pathway enrichment analysis highlighting alterations in adhesion-related pathways. (E–F) Heatmaps of transcription factors (E) and adhesion molecules (F). (G) RNA-seq analysis of genes shown in the figure in BM cells. (H) RNA-seq analysis of genes shown in the figure in human MSCs with or without hIL-27 treatment (n = 2 per group). (I) qPCR analysis of genes shown in the figure in murine MSCs with or without IL-27 (n = 3 per group). (J) RNA-seq analysis of genes shown in the figure in BM Gr-1 + cells 3 weeks after AAV-Ctrl or AAV-IL-27 administration (n = 2 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; unpaired Student's t-test.

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Expressing, Marker, RNA Sequencing

Journal: eBioMedicine

Article Title: Interleukin-27 remodels the bone marrow niche to suppress B-cell development and leukaemia progression in mouse models

doi: 10.1016/j.ebiom.2026.106239

Figure Lengend Snippet: IL-27 exhibits therapeutic potential while modulating B-cell compartments. (A–B) Experimental design of combination therapy in WT (A) and Rag1 −/− (B) mice, with corresponding Kaplan–Meier survival analysis. (C–D) Experimental design of IL-27R −/− CAR-T cell therapy combined with AAV-Ctrl or AAV-IL-27 in B-ALL-bearing mice, with Kaplan–Meier survival curves. B-ALL cells were transplanted either prior to chemotherapy (C) or after chemotherapy (D). (E–F) NSG mice transplanted with human G-PBSCs were injected intramuscularly with AAV-Ctrl or hIL-27. 10 days later, frequencies and absolute numbers of B cells in the BM and spleen were analysed (n = 5 per group). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001; ns, not significant; log-rank test (A–D) and unpaired Student's t-test (E–F).

Article Snippet: Cells were treated with recombinant murine IL-27 (51107-M08H, Sino Biological) or human IL-27 (50 ng/mL) for 24 h prior to RNA extraction.

Techniques: Injection

Journal: International Journal of Molecular Sciences

Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

doi: 10.3390/ijms27073232

Figure Lengend Snippet: Interleukin (IL)−1β levels are elevated in MK2/3 double-knockout (DKO) mice. ( a ) Il1b mRNA levels are increased in untreated and ( b ) IL-1α-treated (5 ng/mL, 1 h) MK2/3 -DKO bone marrow-derived macrophages (BMDMs) compared to wild type (WT). WT n = 14, DKO n = 13. ( c ) Basal IL-1β protein levels are elevated in MK2/3 -DKO BMDM. Elongation factor 2 (EF2) serves as a control. One representative Western blot of total WT n = 4, DKO n = 6. ( d ) The concentration of IL-1β is higher in the supernatant of untreated and ( e ) IL-1α (5 ng/mL, 4 h) + Nigericin (20 µM, 8 h) or ( f ) IL-1α + ATP (5 mM, 5.5 h)-treated MK2/3 -DKO BMDM than in WT. ( d , e ) WT n = 9, DKO n = 8. ( f ) n = 3/group. ( g ) The basal concentration of IL-1β is elevated in the serum of MK2/3 -DKO mice. n = 6 mice/group, whereby one sample/group was pooled from 3 mouse sera. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

Techniques: Double Knockout, Derivative Assay, Control, Western Blot, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

doi: 10.3390/ijms27073232

Figure Lengend Snippet: The level of IL-1β is increased in immortalized MK2 -KO (i MK2 -KO) cells. ( a ) i MK2 -KO cells transduced with an empty vector (+ GFP ) showed elevated levels of Il1b mRNA compared to MK2 -rescued cells (+ MK2 ) (right, n = 8), similar to those observed in MK2/3 -DKO and WT bone-marrow-derived macrophages (BMDMs) (left; WT n = 9, DKO n = 8). ( b ) i MK2 -KO + GFP cells show increased Il1b mRNA after IL-1α treatment (5 ng/mL) compared to MK2 -rescued cells. n = 3. ( c ) RAW cells treated with MK2 siRNA have higher Il1b mRNA levels compared to the control after IL-1α stimulation. ( d ) Similar to MK2 , rescuing MK3 decreases the level of Il1b mRNA in resting or ( e ) IL-1α (5 ng/mL, 1 h)-treated i MK2 -KO cells, ( f ) as well as the basal level of IL-1β protein. Histone H3 serves as a control. ( a , c ) Student’s t -test, ( b ) 2W-RM-ANOVA with Bonferroni posttests, ( d , e ) 1W-ANOVA with Tukey’s Multiple Comparison Test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

Techniques: Transduction, Plasmid Preparation, Derivative Assay, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

doi: 10.3390/ijms27073232

Figure Lengend Snippet: The non-canonical NF-κB pathway is activated in i MK2 -KO cells (mRNA). ( a ) Inhibition of the canonical NF-κB pathway using the IKKβ inhibitors Takinib (10 µM, 2 h) and sc-514 (10 µM, 2 h) reduced Il1b mRNA levels in IL-1α- and LPS-treated i MK2 -KO cells, but did not affect basal Il1b levels. Inhibiting IKKα and IKKβ with HPN-01 (10 µM, 2 h) reduced Il1b mRNA levels in untreated (UT) and IL-1α (5 ng/mL, 1 h)- or LPS-stimulated cells (100 ng/mL, 1 h). Inhibition of the non-canonical NF-κB pathway by IKKα inhibitor B022 (5 µM, 2 h) mainly reduced basal Il1b mRNA. ( b , c ) IL-1β protein level is reduced in resting i MK2 -KO cells after treatment with B022 (5 µM, 7 h). GAPDH serves as a control. ( d ) Il1b mRNA is reduced in MK2/3 -DKO BMDMs after treatment with B022 (5 µM, 2 h). ( e ) The level of Map3k14 mRNA is increased in i MK2 -KO + GFP . ( f ) Relb mRNA level is increased in UT i MK2 -KO + GFP cells. ( g ) Relb and ( h ) Nfkb2 mRNA levels are increased in IL-1α (5 ng/mL, 1 h)-stimulated i MK2 -KO + GFP cells. ( i ) Basal Traf2 mRNA is reduced in i MK2 -KO + GFP cells. ( j ) The Traf3 mRNA level is not changed significantly. ( a ) 1W-ANOVA with Tukey’s Multiple Comparison Test, ( c – j ) Student’s t -test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

Techniques: Inhibition, Control, Comparison

Journal: International Journal of Molecular Sciences

Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

doi: 10.3390/ijms27073232

Figure Lengend Snippet: The non-canonical NF-κB pathway is activated in i MK2 -KO cells (protein). ( a – c ) Compared to i MK2 - KO + MK2 cells, untreated (UT) i MK2 -KO + GFP cells have higher levels of the non-canonical proteins RelB and NF-κB2 in nuclear and cytoplasmic fractions, but not of the canonical RelA and NF-κB1 proteins. Following IL-1α treatment (5 ng/mL, 2 h), the nuclear fraction of i MK2 -KO cells showed elevated protein levels of RelB, NF-κB2, RelA, and NF-κB1. p53 and GAPDH serve as controls for successful nuclear/cytoplasmic separation. EF2 acts as a general control, used for normalization. ( d , e ) The basal protein level of TRAF2 is reduced in whole-cell lysis of i MK2 -KO + GFP cells, whereas TRAF3 and cIAP1/2 are not changed. ( f ) The protein level of c-Rel is increased in i MK2 -KO+ GFP cells. Examples of Western blots from different gels are shown. Mean ± SEM, Student’s t -test, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

Techniques: Control, Lysis, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

doi: 10.3390/ijms27073232

Figure Lengend Snippet: The non-canonical NF-κB pathway is activated in i MK2 -KO cells. RNA sequencing revealed increased levels of mRNA for components of the IL-1β and non-canonical NF-κB pathways, as well as for targets of the non-canonical NF-κB pathway, in i MK2 -KO + GFP cells. This finding was reinforced by IL-1α treatment (5 ng/mL, 1 h).

Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

Techniques: RNA Sequencing

Journal: International Journal of Molecular Sciences

Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

doi: 10.3390/ijms27073232

Figure Lengend Snippet: The MK2 kinase activity is not involved, but the MK2 C-terminus is important. ( a ) The rescued MK2 kinase-inactive mutant, MK2K79R, reduces Il1b mRNA levels to a degree comparable to that of the rescued MK2 in untreated (UT) and ( b ) IL-1α-treated (5 ng/mL, 1 h) i MK2 -KO macrophages. ( c ) i MK2 -KO cells have lower levels of the p38α protein. These levels can be restored by rescuing MK2 , but not by rescuing a MK2 mutant lacking the C-terminus MK2-Δ365–386 . ( d ) MK2-Δ365-386 does not affect the Il1b mRNA levels in IL-1α-treated cells. 1W-ANOVA with Tukey’s Multiple Comparison Test, mean ± SEM, ** p < 0.01, *** p < 0.001.

Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

Techniques: Activity Assay, Mutagenesis, Comparison

Journal: International Journal of Molecular Sciences

Article Title: MK2/p38/p53 Suppress Basal IL-1β and Non-Canonical NF-κB Signaling in Macrophages

doi: 10.3390/ijms27073232

Figure Lengend Snippet: p38α inactivates the non-canonical NF-κB pathway independent of the kinase activity. ( a – c ) Overexpression of p38α in i MK2 -KO cells increases basal TRAF2 and reduces basal RelB protein levels. ( d ) Overexpression of p38α and kinase inactive mutant p38-AGF reduce basal Il1b and ( e ) Map3k14 mRNA and ( f ) increase basal Traf2 mRNA in resting i MK2 -KO cells. ( g ) Relb and ( h ) Nfkb2 mRNA are reduced in IL-1α-treated (5 ng/mL, 1 h) i MK2 -KO+ p38α and +p38-AGF cells. ( b , c ) Student’s t -test, ( d – h ) 1W-ANOVA followed by Tukey’s Multiple Comparison Test, mean ± SEM, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMDM (5 × 10 5 cells/well), i MK2 -KO or RAW 264.7 cells (both 2 × 10 5 cells/well) were seeded, and treated one day later with the indicated concentrations and durations of recombinant murine IL-1α (211-11A, PeproTech GmbH, Hamburg, Germany), LPS ( Escherichia coli 0127:B8; Sigma-Aldrich, Merck, St. Louis, MO, USA), Actinomycin D (Cayman Chemical Company, Ann-Arbor, MI, USA), B022 (MedChemExpress, Monmouth Junction, NJ, USA), BMS-345541 (Axon Medchem B.V., Groningen, Netherlands), CAY10657 (Cayman Chemical Company, Ann-Arbor, MI, USA), SML1160 (Sigma-Aldrich, Merck, St. Louis, MO, USA), IKK inhibitor XII (HPN-01, Sigma-Aldrich, Merck, St. Louis, MO, USA), sc-514 (Cayman Chemical Company, Ann-Arbor, MI, USA), T-5224 (Cayman Chemical Company, Ann-Arbor, MI, USA), Takinib (MedChemExpress, Monmouth Junction, NJ, USA) or Nutlin-3 (SCBT, Dallas, TX, USA).

Techniques: Activity Assay, Over Expression, Mutagenesis, Comparison