Journal: Journal of Translational Medicine

Article Title: TREM-1 triggers necroptosis of macrophages through mTOR-dependent mitochondrial fission during acute lung injury

doi: 10.1186/s12967-023-04027-4

Figure Lengend Snippet: Antibody sources and dilutions

Article Snippet: Then purified macrophages (0.5 × 10 6 cells/well in RPMI 1640) or macrophages pre-treated with the RIPK3 inhibitor (GSK872, 1 μM, Medchem Express, USA), the DRP1 inhibitor (Mdivi-1, 100 nM, Medchem Express) or the mTOR inhibitor (Rapamycin, 100 nM, Beyotime, Jiangsu, China) were added.

Techniques: Western Blot, Immunofluorescence, Flow Cytometry, Staining

Journal: Journal of Translational Medicine

Article Title: TREM-1 triggers necroptosis of macrophages through mTOR-dependent mitochondrial fission during acute lung injury

doi: 10.1186/s12967-023-04027-4

Figure Lengend Snippet: Blockade of TREM-1 reduces macrophage necroptosis in LPS-induced ALI mice. A H&E staining. B the localization of MLKL (green) and F4-80 (red). C – E flow cytometry analysis of MLKL + AlvMs. F , CCK‐8. G – I p-RIPK3 ser232 , RIPK3, p-MLKL ser345 , and MLKL protein. J MLKL (green) expression

Article Snippet: Then purified macrophages (0.5 × 10 6 cells/well in RPMI 1640) or macrophages pre-treated with the RIPK3 inhibitor (GSK872, 1 μM, Medchem Express, USA), the DRP1 inhibitor (Mdivi-1, 100 nM, Medchem Express) or the mTOR inhibitor (Rapamycin, 100 nM, Beyotime, Jiangsu, China) were added.

Techniques: Staining, Flow Cytometry, CCK-8 Assay, Expressing

Journal: Journal of Translational Medicine

Article Title: TREM-1 triggers necroptosis of macrophages through mTOR-dependent mitochondrial fission during acute lung injury

doi: 10.1186/s12967-023-04027-4

Figure Lengend Snippet: TREM-1 activation triggers macrophage necroptosis dependently on RIPK3. A The cell survival rate was examined by CCK‐8, n = 5. B – D p-RIPK3 ser232 , RIPK3, p-MLKL ser345 , and MLKL protein, n = 3. E Immunofluorescence staining for MLKL (green); scale bars = 5 μm. F TNF‐α production in the supernatant. G , H IL-1β p17 level in the supernatant

Article Snippet: Then purified macrophages (0.5 × 10 6 cells/well in RPMI 1640) or macrophages pre-treated with the RIPK3 inhibitor (GSK872, 1 μM, Medchem Express, USA), the DRP1 inhibitor (Mdivi-1, 100 nM, Medchem Express) or the mTOR inhibitor (Rapamycin, 100 nM, Beyotime, Jiangsu, China) were added.

Techniques: Activation Assay, CCK-8 Assay, Immunofluorescence, Staining

Journal: Journal of Translational Medicine

Article Title: TREM-1 triggers necroptosis of macrophages through mTOR-dependent mitochondrial fission during acute lung injury

doi: 10.1186/s12967-023-04027-4

Figure Lengend Snippet: TREM-1 activation induces necroptosis through mitochondrial fission in macrophages. A The cell survival rate, n = 5. B – D Protein levels of RIPK3, MLKL, and their phosphorylation, n = 3. E Immunofluorescence staining for MLKL (green); scale bars = 5 μm. F The concentration of TNF‐α in the supernatant, n = 3. G , H The level of IL-1β p17 in macrophages supernatant, n = 3 biological replicates

Article Snippet: Then purified macrophages (0.5 × 10 6 cells/well in RPMI 1640) or macrophages pre-treated with the RIPK3 inhibitor (GSK872, 1 μM, Medchem Express, USA), the DRP1 inhibitor (Mdivi-1, 100 nM, Medchem Express) or the mTOR inhibitor (Rapamycin, 100 nM, Beyotime, Jiangsu, China) were added.

Techniques: Activation Assay, Immunofluorescence, Staining, Concentration Assay

Journal: Journal of Translational Medicine

Article Title: TREM-1 triggers necroptosis of macrophages through mTOR-dependent mitochondrial fission during acute lung injury

doi: 10.1186/s12967-023-04027-4

Figure Lengend Snippet: mTOR regulates TREM-1-induced necroptosis of macrophages. A the cell survival rate, n = 5. B – D Protein expression of p-RIPK3 ser232 , RIPK3, p-MLKL ser345 , and MLKL, n = 3. E Immunofluorescence staining for MLKL (green) expression; scale bars = 5 μm. F The concentration of TNF‐α was assayed using ELISA, n = 3. G , H IL-1β p17 level in the supernatant, n = 3 biological replicates

Article Snippet: Then purified macrophages (0.5 × 10 6 cells/well in RPMI 1640) or macrophages pre-treated with the RIPK3 inhibitor (GSK872, 1 μM, Medchem Express, USA), the DRP1 inhibitor (Mdivi-1, 100 nM, Medchem Express) or the mTOR inhibitor (Rapamycin, 100 nM, Beyotime, Jiangsu, China) were added.

Techniques: Expressing, Immunofluorescence, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay