Journal: PLoS ONE

Article Title: Fine-Tuning Roles of Endogenous Brain-Derived Neurotrophic Factor, TrkB and Sortilin in Colorectal Cancer Cell Survival

doi: 10.1371/journal.pone.0025097

Figure Lengend Snippet: (A) Role of endogenous BDNF and its receptor TrkB on CRC cell proliferation: effects of exogenous BDNF and suppressing endogenous TrkB receptor on cell proliferation. The four cell lines were cultured for 24 h in FCS-free medium (FCS 10%, −) in the presence of exogenous BDNF (+), K252a (+) alone or in combination. Cell proliferation was determined by flow cytometry analysis using EdU Alexa Fluor 488. The data are presented as histograms of proliferating cells in relative units ± SEM of five independent experiments. *, p <0.05; **, p <0.01; ***, p <0.001, when compared with serum-free medium. (B, C) Effects of exogenous BDNF and suppressing endogenous TrkB receptor on cell survival. Apoptotic ratios of soluble nucleosomes were detected by ELISA Cell for WiDr, SW480, SW620, and COLO 205 induced by serum deprivation alone (FCS 10%, −) or in association either with exogenous BDNF (+), or with K252a (+), during 24–72 h of serum deprivation. Histograms, mean ratio of apoptotic cells ± SEM of at least three independent experiments. *, p <0.05; **, p <0.01; ***, p <0.001, when compared with serum-free condition alone. (D, E) apoptotic ratio after 24 h serum deprivation alone (0% FCS) or with combination with a neutralizing anti-BDNF mAb (0% anti-BDNF). Histograms, mean ratio of apoptotic cells ± SEM of three independent experiments. *, p <0.05; **, p <0.01; ***, p <0.001, when compared with serum-free condition alone.

Article Snippet: Recombinant human BDNF (100 ng/ml), recombinant pro-BDNF (4 ng/ml), and K252a (200 nM) were purchased from Alomone labs.

Techniques: Cell Culture, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Fine-Tuning Roles of Endogenous Brain-Derived Neurotrophic Factor, TrkB and Sortilin in Colorectal Cancer Cell Survival

doi: 10.1371/journal.pone.0025097

Figure Lengend Snippet: (A, B) Sortilin as a coreceptor of p75 NTR . Double staining (yellow) of sortilin (red) and p75 NTR (green) in SW480 cells (A) and SW620 (B) after 24 h of serum deprivation. (C) Colocalization of pro-BDNF and sortilin. Confocal microscopy study of a WiDr cells stained with an anti-pro-BDNF Ab (green) and an anti-sortilin Ab (red), and double staining (yellow) after 24 h of serum deprivation. (D, E) apoptotic ratios after 24 h serum deprivation alone (0% FCS) or combined with recombinant Pro-BDNF (0% Pro-BDNF). Histograms, mean ratio of apoptotic cells ± SEM of three independent experiments. *, p <0.05; **, p <0.01; ***, p <0.001, when compared with serum-free condition alone (0% FCS).

Article Snippet: Recombinant human BDNF (100 ng/ml), recombinant pro-BDNF (4 ng/ml), and K252a (200 nM) were purchased from Alomone labs.

Techniques: Double Staining, Confocal Microscopy, Staining, Recombinant

Journal: Oncotarget

Article Title: miR-1 as a tumor suppressive microRNA targeting TAGLN2 in head and neck squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) The representative quadrant figures of miRNA-control, or miR-1 transfectants in HSC3 [above] and FaDu [below] cells are shown. The bar chart shown on the right of each quadrant indicates the ratio of the early apoptotic cell fraction in miR-1 transfectants in comparison with miRNA-control transfectant. The data for the early apoptotic cell fraction is expressed as the relative value of the average expression of the miRNA-control transfectant. * P < 0.05. (B) The typical figures of cell cycle analysis of miRNA-control, or miR-1 transfectants are shown. The bar chart shown on the right of each figures represents the percentage of the cells in G0/G1, S, or G2/M phase as indicated. * P < 0.05.

Article Snippet: The following RNA species were used in this study: mature miRNAs, Pre-miR™ miRNA Precursors (hsa-miR-1; Pre-miR ID: PM10633), negative control miRNA (P/N: AM17111) (Applied Biosystems, Foster City, CA, USA), small interfering RNA (Stealth Select RNAi™ siRNA; si-TAGLN2 Cat#; HSS144745 and HSS144746) (Invitrogen) and negative control siRNA (D-001810-10; Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Transfection, Expressing, Cell Cycle Assay

Journal: Oncotarget

Article Title: miR-1 as a tumor suppressive microRNA targeting TAGLN2 in head and neck squamous cell carcinoma

doi:

Figure Lengend Snippet: (A) RT-PCR revealed that TAGLN2 mRNA was markedly repressed in miR-1 transfectants compared with the miRNA-controls. * P < 0.05 (B) Western blots revealed that TAGLN2 protein was also decreased in miR-1 .

Article Snippet: The following RNA species were used in this study: mature miRNAs, Pre-miR™ miRNA Precursors (hsa-miR-1; Pre-miR ID: PM10633), negative control miRNA (P/N: AM17111) (Applied Biosystems, Foster City, CA, USA), small interfering RNA (Stealth Select RNAi™ siRNA; si-TAGLN2 Cat#; HSS144745 and HSS144746) (Invitrogen) and negative control siRNA (D-001810-10; Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot