Journal: The Journal of Biological Chemistry

Article Title: Long non-coding RNA LINC00672 contributes to p53 protein-mediated gene suppression and promotes endometrial cancer chemosensitivity

doi: 10.1074/jbc.M116.758508

Figure Lengend Snippet: LINC00672 contributes to the p53-dependent LASP1 suppression. A, association between log2-transformed LINC00672 and LASP1 expression in a set of randomly selected 62 EC tumor and paired adjacent non-neoplastic tissues. B, LINC00672 and LASP1 expression levels detected by qPCR in EC cells after transfection with LINC00672, LINC00672-mutant, and control lentiviruses. Data shown are the mean ± S.D. of three independent experiments, normalized to GAPDH (*, p < 0.05). C, LINC00672 and LASP1 expression levels detected by qPCR in EC cells after transfection with LINC00672 LNA#1, LINC00672 LNA#2 and LNA scramble. Data shown are the mean ± S.D. of three independent experiments, normalized to GAPDH (*, p < 0.05). D, immunofluorescence analysis of LASP1 in EC cells after transfection with LINC00672, LINC00672-mutant, and control lentiviruses. Right panel is the quantification of relative fluorescence mean density (n = 6, *, p < 0.05). E, immunofluorescence analysis of LASP1 in EC cells after transfection with LINC00672 LNA#1, LINC00672 LNA#2, and LNA scramble. Right panel is the quantification of relative fluorescence mean density (n = 6, *, p < 0.05). F, LASP1 levels in HEC-1A cells detected by Western blotting after transfection with LINC00672 lentiviruses, LINC00672-mutant lentiviruses, control lentiviruses, LINC00672 LNA#1, LINC00672 LNA#2, and LNA scramble. G, schema of RNA pulldown experiment followed by Western blotting for verification of hnRNP F/H association (upper). Western blotting results of hnRNP F, H, D, and I in the protein extractions of LINC00672-pulldown are shown. H and I, RIP experiments were performed using hnRNP F, H, D, and I and p53 antibody to immunoprecipitate and a primer to detect LINC00672 in HEC-1A and Ishikawa cells.

Article Snippet: An immunofluorescence analysis of LASP1 was performed using an immunofluorescence staining kit (Beyotime Institute of Biotechnology, China) following the manufacturer's instructions.

Techniques: Transformation Assay, Expressing, Transfection, Mutagenesis, Control, Immunofluorescence, Fluorescence, Western Blot

Journal: eLife

Article Title: Zebrafish fin regeneration involves generic and regeneration-specific osteoblast injury responses

doi: 10.7554/eLife.77614

Figure Lengend Snippet: ( A ) Experimental scheme indicating EdU intraperitoneal (IP) injections used to label cycling osteoblasts harvested at different time points for the data presented in panels B–D. ( B ) Proliferation timeline of bglap :GFP+ osteoblasts in segment –1 as determined by EdU incorporation. N (experiments)=1, n (fins)=4 (0 hpa), 10 (1 day post amputation [dpa]), 9 (2 dpa); n (rays)=14 (0, 2 dpa), 10 (1 dpa). Error bars represent 95% CI. Kruskal-Wallis test. ( C ) Proliferation of bglap :GFP+ osteoblasts in segment –2, segment –1, and segment 0 at 2 dpa. N (experiments)=1, n (fins)=9, n (rays)=14. Error bars represent 95% CI. Dunn’s test. ( D ) bglap RNAscope expression levels of EdU+ cells in segment –1 at 2 dpa. bglap -expressing osteoblasts were categorised based on RNAscope signal intensity into ‘high’ (yellow), ‘medium’ (cyan), and ‘low’ (red) using a look-up table, with ‘high’ threshold corresponding to expression levels in segment –2. N (experiments)=1, n (fins)=9, n (rays)=12, n (cells)=62. Error bars repesent SD. Scale bar, 10 µm.

Article Snippet: To analyse cell proliferation, 5-ethynyl-2′-deoxyuridine (EdU)-Click 647 kit (baseclick GmbH BCK-EdU647) was used.

Techniques: RNAscope, Expressing

Journal: Cancers

Article Title: Heterogeneous Formation of DNA Double-Strand Breaks and Cell-Free DNA in Leukemia T-Cell Line and Human Peripheral Blood Mononuclear Cells in Response to Topoisomerase II Inhibitors

doi: 10.3390/cancers16223798

Figure Lengend Snippet: Time-dependent studies on 53BP1-specific staining in PBMCs and Jurkat cells after treatment with either ETP (25 µM) or DOX (1.0 µM): Automated quantification of cellular immunofluorescence between 0 and 8 h of incubation with topoisomerase II inhibitors using the AKLIDES ® system. DMSO-treated cells were used as a control. Percentage of 53BP1 focus-positive cells in ( A ) PBMCs and ( B ) Jurkat cells were determined at the indicated time points; quantification of 53BP1 foci number per cell in ( C ) PBMCs and ( D ) Jukat cells. Bars represent the mean and standard deviation of four independent experiments. Numbers of asterisks indicate increasing significance levels: * p ≤ 0.05. ETP: etoposide; DOX: doxorubicin.

Article Snippet: The cells were incubated with a commercially provided admixture of two primary antibodies, binding to γH2AX and 53BP1 (Medipan, Germany) in PBS with 1% BSA at the indicated dilutions for 1 h at room temperature.

Techniques: Staining, Immunofluorescence, Incubation, Control, Standard Deviation