t0070907  (MedChemExpress)

Journal: Nature Communications

Article Title: Residual tumor cells after insufficient radiofrequency ablation promote lung metastasis by educating CD177 hi PAD4 hi neutrophils

doi: 10.1038/s41467-025-66897-0

Figure Lengend Snippet: A Conditioned media from normal liver tissues (NLCS), residual tumors post-iRFA (iRFA-TCS), necrotic tissues post-iRFA (iRFA-NCS), and untreated tumors (Ctrl-TCS) were co-cultured with neutrophils. NETs were detected by immunofluorescence. Scale bars, 50 μm. B t-SNE plots comparing Ctrl versus iRFA groups. Ctrl group: 3561 cells; iRFA group: 3453 cells. C GSEA of the PPAR pathway in iRFA-treated vs . untreated CT26 tumors ( n = 3 mice/group). Kolmogorov-Smirnov test. FDR correction. D Lipid metabolite differences between 37 °C-CT26 and 45 °C-CT26 cells ( n = 6 technical replicates). E Tip47 immunofluorescence in liver tumors. Scale bars, 10 μm. F Oil Red O staining of liver tumors (T: tumor, L: liver, N: necrosis). Scale bars, 100 μm and 50 μm. G TG and FA levels in liver (Ctrl-L) and untreated tumors of Ctrl (Ctrl-T), and in the liver (iRFA-L), residual tumors (iRFA-T), and necrotic tissues (iRFA-N) of iRFA (TG: Ctrl-L/Ctrl-T: n = 8, iRFA-L/iRFA-T/iRFA-N: n = 9. FA: Ctrl-L/iRFA-N: n = 11, Ctrl-T: n = 10, iRFA-L: n = 8, iRFA-T: n = 13, mice). P < 0.001, ANOVA. H TG levels in CT26/DLD1 cells incubated at 37 °C/45 °C ± OA ( n = 3 biological replicates). CT26: P < 0.001; DLD1: P = 0.01; ANOVA. I LD area and number in DLD1 cells at 37 °C/45 °C ± OA ( n = 3 biological replicates). P < 0.001 and P = 0.001, respectively; ANOVA. J BODIPY 493 and ( K ) C16 immunofluorescence in cells at 37 °C/45 °C ± OA. Scale bars, 25 μm. L OCR measurements of Basal and Maximal respiration, spare respiration capacity (SRC), and ATP in CT26 cells ( n = 3 biological replicates). Basal, Maximal, and ATP: P < 0.001; SRC: P = 0.001; ANOVA. M FAO rate and ( N ) ROS levels in CT26 cells ( n = 3 biological replicates). M P < 0.001 and (N) P = 0.002, respectively; ANOVA. O , P Immunofluorescence in neutrophils co-cultured with ( O ) OA or ( P ) PF at 37 °C / 45 °C. Scale bars, 50 μm. Data are presented as mean values ± SD. ( O and P ) 37/45: 37 °C/45 °C; TCM: tumor-conditioned media. ( M , N and P ) PF: PF-04620110. Source data are provided as a Source Data file.

Article Snippet: After growing to 60-70% confluence, cells were incubated with OA (OA:10% FBS = 1:10000, 0.31509 mM) (112-80-1, Sigma-Aldrich) for 48 h. DGAT1 inhibtor (25 μM) (PF-04620110, S7192, Selleck), p38 inhibitor (20 μM) (SB202190/SB203580, T2301/T6335, TargetMol), and PPARγ inhibitor (20 μM) (T0070907, S2871, Selleck) were used to inhibit TG formation, p38 signaling pathway, and PPARγ, respectively.

Techniques: Cell Culture, Immunofluorescence, Staining, Incubation

Journal: Nature Communications

Article Title: Residual tumor cells after insufficient radiofrequency ablation promote lung metastasis by educating CD177 hi PAD4 hi neutrophils

doi: 10.1038/s41467-025-66897-0

Figure Lengend Snippet: A PPARa, PPARd, and PPARg mRNA detected via qPCR in cells ( n = 7 biological replicates). ANOVA. B PPRE binding ability for PPAR ( n = 8 biological replicates). ANOVA. C , D Nuclear and cytoplasmic PPARα, PPARβ/δ, and PPARγ proteins in ( C ) tumor cells and ( D ) tumor tissues detected via Western blotting. E , F CD36, FABP4, and FABP5 proteins in ( E ) tumor cells and ( F ) tumor tissues detected via Western blotting. G CD36, FABP4, and FABP5 proteins in CT26 and DLD1 cells treated with siCtrl or si PPARg at 37 °C and 45 °C + OA were detected via Western blotting. H Detection of FA and TG levels in CT26 cells treated with siCtrl or si Pparg at 37 °C and 45 °C + OA (Upper). Detection of FA and TG levels in CT26 cells treated with DMSO or T0 at 37 °C and 45 °C + OA (Lower). FA: n = 3; TG: n = 5; biological replicates. P < 0.001, ANOVA. I Detection of FA and TG levels in DLD1 cells treated with DMSO or T0 at 37 °C and 45 °C + OA (FA: n = 3, TG: n = 5, biological replicates). P < 0.001, ANOVA. J BODIPY C16 was detected in CT26 cells treated with DMSO or T0 at 37 °C and 45 °C + OA via immunofluorescence. Scale bars, 25 μm. K The positive LD area in DLD1 cells treated with DMSO or T0 at 37 °C and 45 °C + OA ( n = 3 biological replicates). P < 0.001, ANOVA. L BODIPY 493 was detected in DLD1 cells treated with DMSO or T0 at 37 °C and 45 °C + OA via immunofluorescence. Scale bars, 25 μm. M ,Nile red staining of CT26 cells treated with siCtrl or si Pparg at 37 °C and 45 °C + OA. Data are presented as mean values ± SD. C , E and G 37/45: 37 °C/45 °C. H – L T0: T0070907. Source data are provided as a Source Data file.

Article Snippet: After growing to 60-70% confluence, cells were incubated with OA (OA:10% FBS = 1:10000, 0.31509 mM) (112-80-1, Sigma-Aldrich) for 48 h. DGAT1 inhibtor (25 μM) (PF-04620110, S7192, Selleck), p38 inhibitor (20 μM) (SB202190/SB203580, T2301/T6335, TargetMol), and PPARγ inhibitor (20 μM) (T0070907, S2871, Selleck) were used to inhibit TG formation, p38 signaling pathway, and PPARγ, respectively.

Techniques: Binding Assay, Western Blot, Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Residual tumor cells after insufficient radiofrequency ablation promote lung metastasis by educating CD177 hi PAD4 hi neutrophils

doi: 10.1038/s41467-025-66897-0

Figure Lengend Snippet: A mTOR, p-mTOR, Akt, p-Akt, P38, and p-P38 proteins detected in CT26 cells at 37 °C/45 °C ± OA. B Representative GSEA analysis of the MAPK pathway in scRNA-seq data from untreated and iRFA-treated CT26 tumors ( n = 3 mice). Kolmogorov-Smirnov test. FDR correction. C P38 and p-P38 in untreated and iRFA-treated tumor tissues. The quantification shown on the right ( n = 9 biological replicates). ANOVA. D Nuclear PPARγ, and cytosol CD36, FABP4, and FABP5 proteins in CT26 cells treated with DMSO or SB202190 at 37 °C and 45 °C + OA. E FA levels in CT26/DLD1 cells treated with DMSO, SB190, or SB580 at 37 °C and 45 °C + OA ( n = 3 biological replicates). P < 0.001, ANOVA. F Nile red staining of CT26 cells treated with DMSO or SB190 at 37 °C and 45 °C + OA. G p-P38 and P38 in CT26 cells treated with siCtrl or si Pparg at 37 °C and 45 °C + OA (Upper), and treated with DMSO or T0070907 (Lower). H p-P38 and P38 proteins in CT26 and DLD1 cells treated with DMSO or PF at 37 °C and 45 °C + OA. I CD36, FABP4, FABP5, PPARγ, and p-P38 staining in liver tumors from iRFA and Ctrl mice. J Pearson correlation between P38 and neutrophil infiltration. K P38 score and neutrophil infiltration score of each patient obtained through ssGSEA. Survival analysis in NEU H p38 H and NEU L wp38 L patients. L MPO-dsDNA levels in neutrophil co-cultured with conditioned media from CT26/DLD1 cells treated with DMSO or SB190 at 37 °C, 37 °C + OA, 45 °C, and 45 °C + OA ( n = 3 biological replicates), P < 0.001, ANOVA. M Lung weight of iRFA and Ctrl CT26 mice treated with SB190 or PBS ( n = 5 mice), P < 0.001, ANOVA. N HE staining of lung tumors. Scale bars, 1 mm and 50 μm. O Immunofluorescence detection of Ly6G and cit-H3 in liver tumors from iRFA-CT26 mice treated with SB190 or PBS. Scale bars, 50 μm. Data are presented as mean values ± SD. A , D , G , and H 37/45: 37 °C/45 °C. E , F , L , M , and O SB190: SB202190. E SB580: SB203580. H PF: PF-04620110. Source data are provided as a Source Data file.

Article Snippet: After growing to 60-70% confluence, cells were incubated with OA (OA:10% FBS = 1:10000, 0.31509 mM) (112-80-1, Sigma-Aldrich) for 48 h. DGAT1 inhibtor (25 μM) (PF-04620110, S7192, Selleck), p38 inhibitor (20 μM) (SB202190/SB203580, T2301/T6335, TargetMol), and PPARγ inhibitor (20 μM) (T0070907, S2871, Selleck) were used to inhibit TG formation, p38 signaling pathway, and PPARγ, respectively.

Techniques: Staining, Cell Culture, Immunofluorescence