Journal: International Journal of Biological Sciences

Article Title: IGF2BP2 Deficiency in Macrophages Impairs Migration, Reprograms Metabolism, and Limits Tumor Progression

doi: 10.7150/ijbs.122142

Figure Lengend Snippet: IGF2BP2 regulates TAM-like polarization and metabolic reprogramming. (A) Gene expression in macrophages from lung adenocarcinoma (TAMs) and adjacent non-tumor tissue (AMs) using publicly available RNA-Seq data from dataset GSE162669 (n = 3 donors per group, triplicates). (B) Gene expression of HMDMs polarized with TCM from A549 cells for 24 h, using publicly available data from GSE162698 (n = 3 individual donors). (C-J) BMMs were cultured in standard medium (M0) or TCM from LLC1 cells (TAM-like) for 8 hours. mRNA expression was measured by qPCR and normalized to Ppia . (C) Igf2bp2 expression is presented as fold change relative to M0 (n = 4 mice per group, duplicates). (D) Expression of TAM-like macrophage markers is shown as a fold change of WT TAM-like (n = 6 mice per group, duplicates). (E, F) ECAR during the Glycolysis Stress Test and (H-J) OCR during the mitochondrial stress test were measured in M0 and TAM-like macrophages using the Seahorse XF Pro Analyzer (n = 4 mice per group, triplicates). (G) mRNA expression of glycolytic markers, expressed as x-fold change relative to TAM-like macrophages (n = 4 mice per group, duplicates). Data are presented as mean ± SEM. Statistical significance was determined by Student's t -test (A-D, G) or ANOVA followed by Bonferroni post-hoc test (F, I, J) (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: A murine cancer model was established by subcutaneous injection of luciferase-expressing Lewis lung carcinoma (LLC1) cells (CRL-1642-LUC2TM, ATCC, Manassas, USA) into 8- to 10-week-old female WT and IGF2BP2 ΜΦ-KO mice.

Techniques: Gene Expression, RNA Sequencing, Cell Culture, Expressing

Journal: International Journal of Biological Sciences

Article Title: IGF2BP2 Deficiency in Macrophages Impairs Migration, Reprograms Metabolism, and Limits Tumor Progression

doi: 10.7150/ijbs.122142

Figure Lengend Snippet: IGF2BP2 deficiency alters membrane lipid composition in macrophages (M0 and TAM-like). BMMs were cultured either in standard medium (M0) or in TCM from LLC1 cells for 8 hours to induce TAM-like polarization. Lipid concentrations in WT and IGF2BP2 KO M0 and TAM-like macrophages were analyzed by mass spectrometry. (A) Hierarchical clustering of log₂-transformed lipid data expressed as a percentage of total lipids, using Ward's linkage and Euclidean distances. Samples are color-coded: WT M0 (dark blue), KO M0 (orange), WT TAM-like (light blue), and KO TAM-like (yellow). (B) PCA of log₂-transformed lipid concentrations (nmol/mg protein) across all lipid species. (C, D) Quantification of free cholesterol (FC) in WT and IGF2BP2 KO macrophages under M0 and TAM-like conditions. (E-H) Concentrations of phosphatidylethanolamine (PE) species (E, G) and PE species grouped by double bond (DB) number (F, H). (I-L) Concentrations of phosphatidylcholine (PC) species (I, K) and PC species grouped by DB number (J, L). Lipid concentrations are expressed as nmol/mg protein. Statistical significance was determined by Student's t-test (C, D) or ANOVA with Bonferroni's post hoc test (E-J). (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: A murine cancer model was established by subcutaneous injection of luciferase-expressing Lewis lung carcinoma (LLC1) cells (CRL-1642-LUC2TM, ATCC, Manassas, USA) into 8- to 10-week-old female WT and IGF2BP2 ΜΦ-KO mice.

Techniques: Membrane, Cell Culture, Mass Spectrometry, Transformation Assay

Journal: International Journal of Biological Sciences

Article Title: IGF2BP2 Deficiency in Macrophages Impairs Migration, Reprograms Metabolism, and Limits Tumor Progression

doi: 10.7150/ijbs.122142

Figure Lengend Snippet: IGF2BP2 ΜΦ-KO reduces tumor growth, alters immune cell infiltration, and impairs angiogenesis in LLC1 tumors. Luciferase-expressing Lewis lung carcinoma (LLC1) cells were injected subcutaneously into WT and IGF2BP2 ΜΦ-KO mice. (A) Tumor volume. (B) Representative images of resected subcutaneous tumors, scale bar = 1 mm per line. (C) Comparison of tumor weights between WT and IGF2BP2 ΜΦ-KO mice. (D) Bioluminescence imaging of luciferase-expressing LLC1 cells following luciferin injection, with photon flux intensity quantified on day 14. (E-J) For flow cytometric analysis, tumors were collected and dissociated on day 14. (E, G, left panel ) Representative dot plots showing TAMs, gated as CD11b⁺F4/80⁺ or CD11b⁺CD68⁺ populations (F, H , right panels ) TAM quantification. (I) TAM polarization. Representative F4/80⁺-gated plots showing CD86 vs. CD206 for WT and KO, plus a merged/overlay panel (WT vs. KO) to illustrate population shifts. (J) Quantification of TAM subsets within F4/80⁺: CD86⁺CD206⁻ (M1-like), CD86⁺CD206⁺ (mixed), and CD86⁻CD206⁺ (M2-like), expressed as % of F4/80⁺ TAMs per tumor. (K, M, O) Representative FSC-A versus CD4, CD8, or NK1.1 plots of viable singlets with a right-hand rectangular gate delineating CD4⁺, CD8⁺, or NK1.1⁺ cell populations. (L, N, P) Quantification of CD4⁺, CD8⁺, and NK1.1⁺ cells, expressed as % of viable singlets per tumor. (Q) Vascularization was quantified as the CD31-positive area fraction. For each field (entire cropped image area), a global threshold was applied to generate a binary red mask, and the %CD31⁺ area was computed as 100 × (area of threshold-positive pixels / total area of the cropped field). Images were acquired with identical exposure settings. Scale bar = 100 µm. ((A-J) n = 8 mice per group, (Q) n = 8 WT, n = 7 ΜΦ-KO mice per group, from two independent experiments). Statistical significance was determined using two-way ANOVA for multiple time points (A) and Student's t -test for comparisons in (C-R) (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: A murine cancer model was established by subcutaneous injection of luciferase-expressing Lewis lung carcinoma (LLC1) cells (CRL-1642-LUC2TM, ATCC, Manassas, USA) into 8- to 10-week-old female WT and IGF2BP2 ΜΦ-KO mice.

Techniques: Luciferase, Expressing, Injection, Comparison, Imaging

Journal: ACS nano

Article Title: Kinetic Control in Assembly of Plasmid DNA/Polycation Complex Nanoparticles

doi: 10.1021/acsnano.9b03334

Figure Lengend Snippet: (A) In vitro transfection efficiencies of nanoparticles (W1–W8, see Table 1) in PC3 cancer cell line (dose = 0.6 μg gWiz-Luc plasmid/104 cells); (B) In vivo transfection efficiency in the lung in healthy BALB/c mice at 12 h post i.v. injection of nanoparticles (W1–W8, see Table 1) containing 40 μg gWiz-Luc plasmid per mouse (left) and representative IVIS images of groups with significant differences in transgene expression (right); (C) In vivo transfection efficiency in the lung of an LL/2 metastasis model in the NSG mice at 48 h post injection of nanoparticles (P1-P8, see Table 1) containing 40 μg PEG-Luc plasmid per mouse (left) and representative IVIS images of groups with significant differences in transgene expression (right); (D) Whole-body biodistributions in BALB/c mice at 1 h post injection of nanoparticles (W1, W2, W6, W8) containing 40 μg 3H-labeled gWiz-Luc plasmid per mouse. Labels: H: heart, K: kidneys, S: stomach, SI: small intestine; (E) Biodistributions to the lung of mice shown in (D); For statistical analysis, n.s. denotes no statistical significance with p > 0.05, *p < 0.05, **p < 0.01, and ***p < 0.001 from one-way or two-way ANOVA and multiple comparisons.

Article Snippet: Preliminary tests revealed that the transgene expression level (luciferase concentration in healthy lung tissues or tumor cells in the lung) peaks at around 12 and 48–72 h post injection for healthy BALB/c mouse model (Jackson Laboratory, US) and LL/2 metastasis in NSG mouse model (Johns Hopkins University Animal Core), respectively.

Techniques: In Vitro, Transfection, Plasmid Preparation, In Vivo, Injection, Expressing, Labeling

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Optimized magnitude of cryosurgery facilitating anti-tumor immunoreaction in a mouse model of Lewis lung cancer

doi: 10.1007/s00262-016-1858-x

Figure Lengend Snippet: a Tumor volume analysis using LL2 cells. Changes of tumor volume of the abscopal tumor. Tumor volume was measured every 2 days. *p < 0.05 between the indicated groups. b Kaplan–Meier plot to show the survival after treatment of the mice in each group. Statistical differences were tested using log-rank test

Article Snippet: Lewis lung carcinoma (LL2) cells and B16 melanoma cells were obtained from DS Pharma Biomedical Corporation (Tokyo, Japan), cultured in DMEM medium supplemented with 15 % fetal bovine serum, and incubated in the specified pathogen-free conditions with 5 % CO 2 at 37 °C.

Techniques: