Journal: Cancer research

Article Title: Stromal platelet-derived growth factor receptor-β signaling promotes breast cancer metastasis in the brain

doi: 10.1158/0008-5472.CAN-19-3731

Figure Lengend Snippet: (A) Pdgfb mRNA is increased in DB7 versus Met1 murine mammary cancer cells (bars represent the mean of biological replicates ± SEM, relative to Gapdh). Western blot confirmation of PDGFB protein expression in DB7 and Met1 cells is shown as an inset. (B) Pdgfrb mRNA is elevated in mouse mammary fibroblasts (MMF) compared to DB7 and Met1 murine mammary cancer cells (bars represent the mean of biological replicates ± SEM, relative to Gapdh). Western blot confirmation of PDGFRβ protein expression in MMF is shown as an inset. (C) Expression of Pdgfb mRNA is diminished with lentiviral transduction of individual shRNA constructs (bars represent the mean of biological replicates ± SEM, relative to Gapdh). Western blot of PDGFB in DB7 shCtrl and shPdgfb cells is shown as an inset. (D) Quantification of orthotopic mammary tumor volume over time [n = 5 DB7 shCtrl, n = 5 DB7 shPdgfb-1, shPdgfb-2 and shPdgfb-3; (**) P = 0.0001 at day 30 by one-way ANOVA]. Error bars represent mean ± SD. (E) Kaplan-Meier survival analysis of DB7 shCtrl and DB7 shPdgfb-3 tumor-bearing mice (n = 5 DB7 shCtrl, n = 4 DB7 shPdgfb-3; P = 0.002). Significance determined by log-rank. (F) Representative images of PDGFB immunostaining (red) on orthotopic mammary tumor tissue (blue = DAPI nuclear counterstain). Scale bars = 20μm.

Article Snippet: DB7 cells were transduced with 3 individual shRNA sequences targeting Pdgfb as well as a scrambled control lentivirus (OriGene Technologies Inc.; #TL501604V).

Techniques: Western Blot, Expressing, Transduction, shRNA, Construct, Immunostaining

Journal: Oncotarget

Article Title: Transcriptional co-activator TAZ sustains proliferation and tumorigenicity of neuroblastoma by targeting CTGF and PDGF-β

doi:

Figure Lengend Snippet: (A) BE(2)-C cells were overexpressed with TAZ or overexpressed TAZ cells were knocked down with TAZ siRNA. Total cellular extracts were prepared and subjected to Western blot using antibodies against CTGF and PDGF-β. (B) SK-N-AS cells were knocked down with TAZ siRNA or TAZ siRNA cells were re-introduced with TAZ. Total cellular extracts were prepared and subjected to Western blot using antibodies against CTGF and PDGF-β. GAPDH levels are shown as loading control.

Article Snippet: Mouse anti-TAZ (560235; BD Biosciences), mouse anti-YAP (sc-101199, 1:200), goat anti-CTGF (sc-14939, 1:200) from Santa Cruz company, rabbit anti-PDGF-β (E1A0240-1, 1:1000) from EnoGene, and mouse anti-GAPDH (AG019, 1:1000) from Beyotime, cell cycle regulation antibody sampler kit #9932 from Cell Signaling Technology were used as primary antibodies.

Techniques: Western Blot

Journal: Oncotarget

Article Title: Transcriptional co-activator TAZ sustains proliferation and tumorigenicity of neuroblastoma by targeting CTGF and PDGF-β

doi:

Figure Lengend Snippet: TAZ overexpressing BE(2)-C cells were infected with PDGF-β siRNA. (A) qRT-PCR analysis was performed to determine mRNA expression of PDGF-β. Data represent the means ± SD from three independent experiments. (* P < 0.05). (B) Western blot analysis was performed to determine PDGF-β expression. GAPDH levels are shown as loading control. (C) Cells were seed into a 96-well plate (1000 cells/well), and cell proliferation was determined using cell counting kit-8 assay kit. Data represent the means ± SD from three independent experiments. (* P < 0.05). (D) Colony formation was examined by soft agar assay. (E) Colony number was counted using counter. The values represent the mean ± SD from three independent experiments (* P < 0.05). (F) Cell cycle was analyzed using PI staining and flow cytometry. (G) Immunoblot analysis was performed to determine the expression of G1 cell cycle regulatory proteins including cyclin D1, CDK4 and CDK6. GAPDH levels are shown as loading control.

Article Snippet: Mouse anti-TAZ (560235; BD Biosciences), mouse anti-YAP (sc-101199, 1:200), goat anti-CTGF (sc-14939, 1:200) from Santa Cruz company, rabbit anti-PDGF-β (E1A0240-1, 1:1000) from EnoGene, and mouse anti-GAPDH (AG019, 1:1000) from Beyotime, cell cycle regulation antibody sampler kit #9932 from Cell Signaling Technology were used as primary antibodies.

Techniques: Infection, Quantitative RT-PCR, Expressing, Western Blot, Cell Counting, Soft Agar Assay, Staining, Flow Cytometry