Journal: Journal of Translational Medicine

Article Title: Induction of filopodia formation by α-Actinin-2 via RelA with a feedforward activation loop promoting overt bone marrow metastasis of gastric cancer

doi: 10.1186/s12967-023-04156-w

Figure Lengend Snippet: ACTN2 is a direct target gene of the NF-κB signaling in GC cells. A c-Fos, NFATc1, RelA, Smad3 or Smad4 was cotransfacted with the ACTN2 promoter (-1918/ + 2455) reporter gene ( ACTN2 Luc-R) into SNU-16 cells for 12 h and dual-luciferase reporter assays were performed. B ACTN2 promoter 5′ sequential deletion constructs. Fragments of different lengths of the ACTN2 promoter with the same 3′ end were cloned into pGL3-Basic. RelA was co-transfected with each of the ACTN2 promoter-based reporters for 12 h and dual-luciferase reporter assays were performed. C Mutation of putative RelA binding site at the ACTN2 promoter. SNU-16 cells were transfected with wild type or mutant ACTN2 Luc-R for 12 h and dual-luciferase reporter assays were performed. The statistical analysis information in A-C: The bars indicate the SD. The results are expressed as the mean ± SD of three independent experiments. * P < 0.05 using Ordinary one-way ANOVA. D NAGC and HAGC tissue sections obtained after infusion were immunostained for α-Actinin-2 (green), Ser276 phosphorylated RelA (p-RelA, red) and nucleus (blue). E Differences in expression of α-Actinin-2 and p-RelA between HAGC and NAGC, and relationship of overall survival proportion of patients based on univariate analyses. * P < 0.05 using the two-sided Pearson chi-squared tests. F Overall survival proportion of patients with HAGC and NAGC stratified by α-Actinin-2 and p-RelA simultaneous high expression based on Kaplan–Meier survival analyses. HAGC-yes: α-Actinin-2 and p-RelA were simultaneously highly expressed in HAGC sample. HAGC-no: at least one marker in α-Actinin-2 and p-RelA had low expression in HAGC sample. Same to NAGC-yes and NAGC-no. * P < 0.05 using Kaplan–Meier plots and compared with the log-rank test. G and H SUN-16 cells were transfected with or without RelA for 24 h, then the cell nuclear extracts were subjected to ChIP analysis using the p-RelA antibody. G Western blotting analysis with the p-RelA antibody demonstrates the IP specificity and efficiency. H DNA isolated and purified from immunoprecipitated material was amplified with primers spanning the RelA binding site (Fs/Rs) or primers far away from the RelA binding site (Ff/Rf) of the ACTN2 gene promoter

Article Snippet: * P < 0.05 using the two-sided Pearson chi-squared tests.

Techniques: Luciferase, Construct, Clone Assay, Transfection, Mutagenesis, Binding Assay, Expressing, Marker, Western Blot, Isolation, Purification, Immunoprecipitation, Amplification

Journal: Nature Communications

Article Title: Clinical features associated with Neo RAS wild-type metastatic colorectal cancer A SCRUM-Japan GOZILA substudy

doi: 10.1038/s41467-024-50026-4

Figure Lengend Snippet: Forest plot depicting the logistic regression multivariate analysis for the appearance of Neo RAS WT mCRC in Group A (4a) and Group B (4b). The adjusted odds ratio (OR: blue plot) and 95% confidence interval (CI: black horizontal lines) are shown for each factor. Various clinical factors and their association with the appearance of Neo RAS WT mCRC, as indicated by OR, are analyzed across the cohort using the two-sided Wald chi-squared test. The unadjusted OR of the appearance of Neo RAS WT mCRC is calculated as X/Y. For instance, about liver metastasis, X is defined as “the proportion of mCRC with liver metastases that became Neo RAS WT mCRC,” and Y is defined as “the proportion of mCRC without liver metastases that became Neo RAS WT mCRC.” An adjusted OR is an OR that has been adjusted to account for other predictor variables in the model (adjusted OR: 0.18, 95% CI: 0.09–0.34, P = 1.43 × 10 −7 ). Vertical dotted line: the null hypothesis. Number of events: Group A = 91; Group B = 42. Source data are provided as a Source Data file. mCRC metastatic colorectal cancer, WT wild-type, RAS rat sarcoma viral oncogene homolog, VEGF vascular endothelial growth factor.

Article Snippet: Various clinical factors and their association with the appearance of Neo RAS WT mCRC, as indicated by OR, are analyzed across the cohort using the two-sided Wald chi-squared test.

Techniques: