Journal: Cell

Article Title: Oncogenic Role of THOR , a Conserved Cancer/Testis Long Noncoding RNA

doi: 10.1016/j.cell.2017.11.040

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: IGF2 cDNA , Origene , SC119786.

Techniques: Virus, Plasmid Preparation, Transgenic Assay, Recombinant, TA Cloning, Immunoprecipitation, Software, CRISPR

Journal: Advanced Biology

Article Title: LIN28B Promotes Cancer Cell Dissemination and Angiogenesis

doi: 10.1002/adbi.202400730

Figure Lengend Snippet: LIN28B promotes angiogenesis through IGF2 secretion. A) The level of IGFs and IGFRs of CTRL and iLIN28B‐derived conditioned media was determined using a Quantibody human IGF signaling array Q1. Fold change values were compared to CTRL (CTRL = 1, dashed line). B) Immunoblot analysis for IGF2 on CTRL and iLIN28B CM. Coomassie‐brilliant‐blue staining was shown as loading control. The molecular weights are indicated in KiloDaltons (KDa). Densitometric analysis of Western blots was performed by ImageJ software and the results were represented as fold change with respect to CTRL CM. C) Real‐time qPCR analysis for total IGF2 mRNA content in CTRL and iLIN28B cells. The expression of GAPDH was used for data normalization. D) Real‐time qPCR analysis of the indicated nascent transcripts in CTRL and iLIN28B cells. The expression of GAPDH was used for normalization of each sample. E) The binding of LIN28B with the indicated mRNAs was tested by RIP analysis through real‐time qPCR analysis. Data were then plotted as the fold enrichment of mRNAs in the iLIN28B immunoprecipitated (IP) sample relative to the ones observed in the IgG IP samples. Bottom: the presence of LIN28B in the IP material was confirmed by Immunoblot analysis. F) Top: representative brightfield images of the vessel‐like structures formed by endothelial cells in the presence of iLIN28B and CTRL CM pre‐incubated with IGF2 blocking antibody (IGF2 Ab). Where indicated, endothelial cells were pre‐treated with PPP. Scale bar, 300 µm. Bottom: quantification of meshes and nodes. G) Left: confocal Z‐stack reconstructions of Tg(KDRL:GFP) zebrafish embryos engrafted with CTRL, iLIN28B, and iLIN28B (red) resuspended with IGF2 blocking antibody (IGF2 Ab). For vessel quantification, the embryos were stained for the AP; n = 30. Asterisks indicate the site of engraftment. Scale bars, 100 µm. Right: quantification of the mean number of vessels from the SIV towards the tumor mass and within the SIV. Data are presented as the mean number ± standard error (S.E.M.). * P < 0.05; ** P < 0.01 compared to CTRLs (Student's t ‐test).

Article Snippet: For neutralization experiments, CM was preincubated with the antibody against human IGF2 (0.8 μg mL −1 ; AF‐292‐NA; R&D Systems, Minneapolis, MN, USA) overnight at 4 °C.

Techniques: Derivative Assay, Western Blot, Staining, Control, Software, Expressing, Binding Assay, Immunoprecipitation, Incubation, Blocking Assay