Journal: Clinical Epigenetics

Article Title: Comprehensive analyses of molecular features, prognostic values, and regulatory functionalities of m 6 A-modified long non-coding RNAs in lung adenocarcinoma

doi: 10.1186/s13148-023-01475-z

Figure Lengend Snippet: The expression levels of m 6 A regulators in LUAD and the correlation between m 6 A regulators. A Heatmap of the expression levels of m 6 A regulators (LUAD tumor tissues vs. normal lung tissues) in TCGA and GTEx databases. FC represented the expression fold change of T-median/N-median. WilcoxTest. * P < 0.05, ** P < 0.01, *** P < 0.001 and not significant (ns) P > 0.05. B RT-qPCR showed the relative mRNA expression levels of METTL3, METTL14, ALKBH5, FTO, and IGF2BP1/3 in eight paired LUAD tissues (Tumor) and adjacent non-tumor tissues (Normal). Paired t test. ** P < 0.01 and not significant (ns) P > 0.05. C, D Western blots showed the protein expression of METTL3, METTL14, FTO, ALKBH5, and IGF2BP1/3 in LUAD tissues ( C ) and cell lines ( D ), respectively. E The Pearson correlation analysis of m 6 A regulators’ expression levels in 872 samples in TCGA and GTEx databases. The number in the circle represented the Pearson coefficient, which determined the size of circle. The cross in the circle represented no correlation ( P > 0.001). F The PPI network analysis of m 6 A regulators. Line thickness indicated the strength of data support

Article Snippet: The primary antibodies were as follows: β-actin (4970S, 1:1000, Cell Signaling Technology (CST), Beverly, MA, USA), METTL3 (86132S, 1:2000, CST, Beverly, MA, USA), METTL14 (26158-1-AP, 1:2000, Proteintech, Wuhan, China), ALKBH5 (80283S, 1:1000, CST, Beverly, MA, USA), FTO (27226-1-AP, 1:2000, Proteintech, Wuhan, China), IGF2BP1 (EPR26408-18, 1:2000, Abcam, Cambridge, UK), and IGF2BP3 (14642-1-AP, 1:2000, Proteintech, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Molecular Oncology

Article Title: Pericyte‐secreted IGF2 promotes breast cancer brain metastasis formation

doi: 10.1002/1878-0261.12752

Figure Lengend Snippet: Insulin‐like growth factor expression in pericytes. (A, B) Igf1 and Igf2 mRNA expression in human and mouse brain and tumor cells, as assessed by qPCR with GAPDH as housekeeping gene. Results are shown as fold change in comparison with Igf1 in pericytes. Inset in (A) shows human Igf1 expression in a lower scale range. N = 3, average ± SD (statistically not analyzed). (C) IGF2 secretion in human brain pericytes and astrocytes, as assessed by ELISA. N = 3 pericytes, N = 2 astrocytes, average ± SD, * P < 0.05, ** P < 0.01 (Student's t ‐test). (D) Representative immunofluorescence staining showing co‐localization of IGF1 and CD13 in human TNBC brain metastatic tissue. Scale bar = 10 µm. (E) Representative immunofluorescence staining showing co‐localization of IGF2 and CD13 in human TNBC brain metastatic tissue. Scale bar = 10 µm.

Article Snippet: Insulin‐like growth factor 1 (IGF1) and IGF2 contents of conditioned media were measured using commercial ELISA kits (CSB‐E04580h and CSB‐E04583h, respectively; Cusabio, Wuhan, China) following the manufacturer's instructions.

Techniques: Expressing, Comparison, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

Journal: Molecular Oncology

Article Title: Pericyte‐secreted IGF2 promotes breast cancer brain metastasis formation

doi: 10.1002/1878-0261.12752

Figure Lengend Snippet: Role of IGFs in breast cancer cell proliferation in vitro . (A) Quantification of MDA cell growth in control or pericyte‐conditioned media in the presence or absence of PPP. Cells were counted in phase‐contrast micrographs. N = 5, average ± SD, * P < 0.05 (ANOVA and Bonferroni's post hoc test). (B) Growth of 4T1 cells in control or pericyte‐conditioned media in the presence or absence of PPP, as assessed by impedance measurements. N = 4, average ± SD, P < 0.001 (cells cultured in pericyte‐conditioned media compared to control), P < 0.05 (cells treated with PPP compared to control; cells cultured in pericyte‐conditioned media and PPP compared to cells cultured in pericyte‐conditioned media) (two‐way ANOVA with repeated measures and Tukey's post hoc test). (C) Growth of MDA cells cultured in control or pericyte‐conditioned media in the presence or absence of PPP, or in conditioned media of Lipofectamine‐treated or Igf2‐silenced pericytes. N = 5, average ± SD, ** P < 0.01 (compared to control), ## P < 0.01 (compared to cells cultured in pericyte‐conditioned media) (ANOVA and Bonferroni's post hoc test). (D) Effect of Igf2 silencing on Igf2 and Igf1 mRNA expression in HBVP cells, as assessed by qPCR with GAPDH as housekeeping gene. N = 3, average ± SD, P < 0.05 (ANOVA and Bonferroni's post hoc test). (E) Igf1R and Igf2R mRNA expression in human tumor cells. Results are shown as fold change in comparison with Igf1R in breast cancer cells. N = 3, average ± SD, * P < 0.05 (ANOVA and Bonferroni's post hoc test).

Article Snippet: Insulin‐like growth factor 1 (IGF1) and IGF2 contents of conditioned media were measured using commercial ELISA kits (CSB‐E04580h and CSB‐E04583h, respectively; Cusabio, Wuhan, China) following the manufacturer's instructions.

Techniques: In Vitro, Control, Cell Culture, Expressing, Comparison