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. 2024 Mar 7;9:64. doi: 10.1038/s41392-024-01772-w

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — CPT1A-mediated ferroptosis resistance in LCSCs relies on TAMs-derived L-carnitine. a Lipid ROS levels in LLC-shNC/shCPT1A cells treated with L-carnitine (10 mM) or not for 2 days and then erastin (2 μM) for 6 h determined by flow cytometry. Data represent the mean ± s.d.; n = 2 samples. Effects of L-carnitine (IG, 100 mg/kg) on tumor progression in tumor-bearing mice inoculated with LLC-shNC/shCPT1A cells. Tumor growth curve (data represent the mean ± s.e.m.; n = 5 samples) (b). Representative images of C11-BODIPY staining in the tumor. Scale bar: 50 μm (insets) (c). Percentages of CD8⁺ or CD69⁺CD8⁺ cells in the tumor. Data represent the mean ± s.d.; n = 3-5 samples (d). e L-carnitine levels in the serum of Kras mutant or wild type mice. measured by colorimetry. Data represent the mean ± s.d.; n = 5 samples.L-carnitine levels in the virous cell types of tumor-bearing mice measured by colorimetry. L-carnitine levels in CD45^-/CD11b⁺/CD3⁺ cells isolated from the tumor (f) and in macrophages isolated from the spleen or the tumor (g) in mice. Data represent the mean ± s.d.; n = 3 samples. h L-carnitine levels of intracellular or extracellular of M0/M1/M2-macrophages measured by colorimetry. Data represent the mean ± s.d.; n = 3 samples. i Heatmap of metabolites up-regulated in the THP-1-M2 medium calculated by LC-MS (P < 0.05; one-way ANOVA) (n = 5 samples). j Representative western blot for BBOX1 and OCTN2 in RAW264.7-M0/M1/M2 cells or LLC cells (left). Representative western blot for OCTN2 in CD44^+/- cells or ALDH^+/- cells of LLC cells (right). k Absorbed L-carnitine levels in CD44^+/− cells treated with L-carnitine or not for 2 days measured by colorimetry. Data represent the mean ± s.d.; n = 5 samples. l Lipid ROS levels in LLC cells co-cultured with Raw264.7-M2 cells (treated with mildronate or not) or not determined by flow cytometry. Data represent the mean ± s.d.; n = 3 samples. m Overall survival of NSCLC patients stratified according to tumor BBOX1 or OCTN2 expression analyzed with Kaplan-Meier Plotter websites. Research model of CPT1A/c-Myc loop regulating ferroptosis in LCSCs. *P < 0.05, **P < 0.01, ***P < 0.001; ns: not significant. The statistical analysis was performed using a two-tailed Student’s t-test or Pearson’s correlation test