Journal: Cell Research
Article Title: Dynamic magneto-mechanical force in lysosomes induces durable macrophage repolarization for antitumor immunity
doi: 10.1038/s41422-025-01217-1
Figure Lengend Snippet: a RAW 264.7 cells were polarized into M0, M1 (induced by IFN-γ and LPS), or M2 (induced by IL-4 and IL-13) macrophages. M0 macrophages were incubated with MNMs and then treated with 1 Hz RMF for 15 min. Macrophages were stained with ActinRed (red), and nuclei were stained with Hoechst (blue). b Polarized M0 RAW 264.7 cells were incubated with MNMs and then treated with 1 Hz RMF for 15 min. Flow cytometry analysis of CD86 and CD206 expression was performed. c , d RNA-sequencing analyses of polarized M2 RAW 264.7 cells treated with or without 1 Hz RMF for 15 min. KEGG enrichment analysis and heat map analysis were performed ( n = 3 independent biological replicates). e , f RAW 264.7 cells or BMDMs were polarized into M0, M1, or M2 macrophages. M2 macrophages were incubated with MNMs and then treated with or without 1 Hz RMF for 15 min (M2-MNM represents M2 macrophages incubated with MNMs, and M2-RMF represents M2 macrophages incubated with MNMs and treated with RMF). Concentrations of M1-associated proteins (IL-1β) and M2-associated proteins (IL-10) were examined in the conditioned medium of these cells. Data are presented as means ± SD. Statistical significance was defined as P < 0.05 ( n = 3 independent biological replicates). g RAW 264.7 cells were polarized into M0, M1, or M2 macrophages. M0 macrophages were cultured in conditioned medium derived from LLC and incubated with or without MNMs, then treated with or without 1 Hz RMF for 15 min. mRNA levels of M1-associated genes ( Cd80 , Cd86 , Il1b , Il6 ) and M2-associated genes ( Cd163 , Cd206 , Il4 , Il10 ) were examined. Data are presented as means ± SD ( n = 4 independent biological replicates).
Article Snippet: To explore the proportions of different macrophage subtypes in tumor tissues after MagLMP treatment, macrophages in the sections were stained by immunofluorescence with anti-mouse CD206 (R&D Systems, AF2535), anti-mouse CD80 (Proteintech, 14292-1-AP), anti-mouse F4/80 (R&D Systems, MAB5580), and DAPI (Beyotime Biotech, C1002).
Techniques: Incubation, Staining, Flow Cytometry, Expressing, RNA Sequencing, Cell Culture, Derivative Assay
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