Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet: Lung cancer cells induced CAFs activation via IGF2 secretion (A) mRNA levels of IGF2 in cells were detected by qPCR. (B) IGF2 concentrations in cell culture medium were detected by ELISA. (C) NFs were treated with IGF2 (50 and 100 ng/mL) for 24 h. Cell morphology was observed under a microscope. (D) NFs were treated with IGF2. α-SMA and FAP expressions were detected by western blotting. (E) NFs were treated with IGF2. Cell proliferation was examined by CCK-8 kit after 48 h. (F and G) NFs were treated with IGF2. NFs migration was detected by trans-well assay after 24 h. (H) NFs were treated with CM or CM + IGF2 neutralizing antibody. Cell morphology was observed under a microscope. (I) NFs were treated with CM or CM + IGF2 neutralizing antibody. α-SMA and FAP expressions were detected by western blotting. (J) NFs were treated with CM or CM + IGF2 neutralizing antibody, and IGF2 (100 ng/mL). Cell proliferation was examined by CCK-8 kit after 48 h. (K and L) NFs were treated with CM or CM + IGF2 neutralizing antibody. NFs migration was detected by trans-well assay after 24 h. (M) The effect of IGF2 knockdown by RNAi. (N) IGF2 expression was knocked down in lung cancer cells, and then the CM was collected and used to culture NFs. α-SMA and FAP expressions were detected by western blotting. (O and P) IGF2 expression was knocked down in lung cancer cells by RNAi, and then the CM was collected and used to culture NFs. NFs migration was detected by trans-well assay after 24 h. Ab, neutralizing antibody. Scale bar, 100 μm. Data represents the mean ± SD from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 .

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques: Activation Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Microscopy, Western Blot, CCK-8 Assay, Migration, Knockdown, Expressing

Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet: IGF2-induced autophagy mediated the effect of lung cancer cells on CAFs activation (A) NFs were treated with CM and then were stained with 0.5 μg/mL AO for 15 min. The acidic vesicular organelles (AVOs) formation was observed under a fluorescence microscope (scale bar, 20 μm). (B) NFs were treated with CM. The expression of LC-3II and ATG5 were detected by western blotting. (C) NFs were treated with IGF2 (50 or 100 mg/mL) for 24 h. The AVOs formation was examined by AO staining (scale bar, 20 μm). (D) NFs were treated with IGF2. The expression of LC-3II and ATG5 was detected by western blotting. (E) NFs were treated with IGF2 or IGF2 + 3-MA (5 mM). The AVOs formation was examined by AO staining (scale bar, 20 μm). (F) NFs were treated with IGF2 or IGF2 + 3-MA. Protein expressions were detected by western blotting. (G) NFs were treated with IGF2 or IGF2 + 3-MA. Cell proliferation was examined by CCK-8 kit after 48 h. (H and I) NFs were treated with IGF2 or IGF2+3-MA. NFs migration was detected by trans-well assay after 24 h (scale bar, 100 μm). (J) NFs were treated with IGF2, CM, CM + IGF2 Ab, and CM + 3-MA. The AVOs formation was examined by AO staining (scale bar, 20 μm). (K) NFs were treated with CM, IGF2, and CM + IGF2 Ab. Protein expressions were detected by western blotting. (L) NFs were treated with IGF2, CM, CM + IGF2 Ab, and CM + 3-MA. Cell proliferation was examined by CCK-8 kit after 48 h. (M and N) NFs were treated with IGF2, CM, CM + IGF2 Ab, and CM + 3-MA. NFs migration was detected by trans-well assay after 24 h (scale bar, 100 μm). (O and P) NFs were treated with RAPA (100 nM); α-SMA and FAP expressions were detected by western blotting (scale bar, 20 μm). (Q and R) NFs were treated with RAPA; NFs migration was detected by trans-well assay after 24 h. Ab, neutralizing antibody (scale bar, 100 μm). Data represents the mean ± SD from three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 .

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques: Activation Assay, Staining, Fluorescence, Microscopy, Expressing, Western Blot, CCK-8 Assay, Migration

Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet:

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Reprogramming of fibroblasts into cancer-associated fibroblasts via IGF2-mediated autophagy promotes metastasis of lung cancer cells

doi: 10.1016/j.isci.2024.111269

Figure Lengend Snippet: PCR primer sequences

Article Snippet: Recombinant human IGF2 , Sino Biological , Cat#13032-HNAY.

Techniques:

Journal: Discover Oncology

Article Title: Novel N 6 -methyladenosine (m 6 A) writer METTL16 promotes the cervical cancer tumorigenesis by targeting FTH1-dependent ferroptosis

doi: 10.1007/s12672-026-04403-8

Figure Lengend Snippet: FTH1 acted as the target of METTL16. A The immunofluorescence assay showed the subcellular localization of METTL16 and FTH1. B The binding motif sites of METTL16 for its target genes (GGAAC). C The FTH1 level in cervical cancer cells was test by the RT-PCR. D RNA immunoprecipitation (RIP) assay was conducted for the binding within METTL16 and FTH1 in HeLa cells. E RIP assay using anti-m 6 A conducted for the m 6 A modification on FTH1 mRNA. * p < 0.05, ** p < 0.01

Article Snippet: Cervical cancer cells were lysed by immunoprecipitation buffer (protease, phosphatase inhibitors, MCE) containing magnetic beads conjugated to specific antibodies (anti-IGF2BP2, anti-METTL16, anti-m 6 A).

Techniques: Immunofluorescence, Binding Assay, Reverse Transcription Polymerase Chain Reaction, RNA Immunoprecipitation, Modification

Journal: Discover Oncology

Article Title: Novel N 6 -methyladenosine (m 6 A) writer METTL16 promotes the cervical cancer tumorigenesis by targeting FTH1-dependent ferroptosis

doi: 10.1007/s12672-026-04403-8

Figure Lengend Snippet: IGF2BP2 promoted the stability of FTH1 mRNA. A The immunofluorescent staining showed the subcellular localization of IGF2BP2 and FTH1 in HeLa cells. B The positive correlation within IGF2BP2 and FTH1 in clinical samples ( http://gepia.cancer-pku.cn/index.html ). C RNA immunoprecipitation (RIP) assay showed the immunoprecipitated FTH1 mRNA by anti-IGF2BP2 antibody. D RNA decay analysis reported the stability of FTH1 mRNA in cervical cancer cells with IGF2BP2 overexpression. E RNA decay analysis reported the stability of FTH1 mRNA in cervical cancer cells with METTL16 silencing (sh-METTL16-1, sh-METTL16-2). F RNA decay analysis reported the stability of FTH1 mRNA in cervical cancer cells with IGF2BP2 silencing (si-IGF2BP2) and METTL16 overexpression. * p < 0.05, ** p < 0.01

Article Snippet: Cervical cancer cells were lysed by immunoprecipitation buffer (protease, phosphatase inhibitors, MCE) containing magnetic beads conjugated to specific antibodies (anti-IGF2BP2, anti-METTL16, anti-m 6 A).

Techniques: Staining, RNA Immunoprecipitation, Immunoprecipitation, Over Expression