Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma-5

<p><b>Copyright information:</b></p><p>Taken from "Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma"</p><p>http://www.biomedcentral.com/1471-2407/7/224</p><p>BMC Cancer 2007;7():224-224.</p><p>Published online 12 Dec 2007</p><p>PMCID:PMC2234427.</p><p></p>e subcutaneously injected on both sides of the midline of the lower dorsal abdomen of wild type (wt) or D849N-mutant mice (mut). Six tumors were analyzed for each experimental combination. When the B16 melanomas had reached 0.8 cm, they were removed and analyzed for ratio of total vessel area/viable tumor (A), average vessel surface (B), number of vessels per high-power field (C) and numerical density of pericytes per perimeter of vessel (D). Asterisks indicate statistical significance (*, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001) by Student's -test

Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma-6

<p><b>Copyright information:</b></p><p>Taken from "Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma"</p><p>http://www.biomedcentral.com/1471-2407/7/224</p><p>BMC Cancer 2007;7():224-224.</p><p>Published online 12 Dec 2007</p><p>PMCID:PMC2234427.</p><p></p>gure 1. Double immunohistochemical staining for endothelial cells (CD31, blue) and perivascular cells (α-smooth muscle actin, brown) in B16 melanoma is shown. Please note the small vessels (blue) covered by few pericyte (brown) in tumors grown in wild type mice (A) compared to larger vessels surrounded by multiple perivascular cells (brown) in tumors grown in D849N-mutant mice (B). Tissue sections were analyzed for total vessel area (C), average vessel surface (D), number of vessels per high-power field (E) and numerical density of pericytes per vessel perimeter (F). Asterisk indicates statistical significance (*, p < 0.01) by Student's -test

Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma-3

<p><b>Copyright information:</b></p><p>Taken from "Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma"</p><p>http://www.biomedcentral.com/1471-2407/7/224</p><p>BMC Cancer 2007;7():224-224.</p><p>Published online 12 Dec 2007</p><p>PMCID:PMC2234427.</p><p></p>bed in the legend of figure 2. Tissue sections were analyzed for the ratio of total vessel area/tumor (A), average vessel surface (B), number of vessels per high-power field (C) and numerical density of pericytes per perimeter of vessel (D). Asterisks indicate statistical significance (*, p < 0.01) by Student's -test. Double immunohistochemical staining for vessels (CD31, blue) and perivascular cells (alpha-smooth muscle actin, brown) identify significantly larger vessels in B16 melanoma/matrigel plugs grown in wild type mice (E) compared to D849N-mutant mice (F) (the same magnification was used in Fig. 4E and Fig. 4F)

Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma-2

<p><b>Copyright information:</b></p><p>Taken from "Platelet-derived growth factor receptor-β, carrying the activating mutation D849N, accelerates the establishment of B16 melanoma"</p><p>http://www.biomedcentral.com/1471-2407/7/224</p><p>BMC Cancer 2007;7():224-224.</p><p>Published online 12 Dec 2007</p><p>PMCID:PMC2234427.</p><p></p> Asterisk indicates statistical significance (*, p < 0.05) by Student's -test

NR4A1 promotes PDGF-BB-induced anchorage-independent growth.

<p>NR4A1 was overexpressed by stable transfection in NIH3T3 cells (A), and U-251MG (B) and U-105MG (C) glioblastoma cells; cells were cultured in soft agar in starvation medium (3% serum) with or without 50 ng/ml PDGF-BB. The number and size of the colony forming unit (CFU) were measured after 10 days. For each cell line a representative experiment is shown, out of at least two independently performed, and the standard deviation between three replicates is indicated in the error bar. A p-value≤0.05 is indicated with an asterisk (*).</p

PDGF-BB induces NR4A1 via Erk1/2 and Erk5.

<p>NIH3T3 cells were serum-starved overnight in 0.1% BSA and treated with inhibitors starting 1 h before stimulation with PDGF-BB (20 ng/ml) for the indicated time periods. Total cell lysate (TCL) (A and C) were analyzed by immunoblotting (Ib) using NR4A1, Erk2, phospho-Erk1/2 and Erk5 antibodies. NR4A1 mRNA was measured with quantitative RT-PCR and panel B shows one out of three independent experiments performed; error bars indicate the standard deviation between three replicates. An asterisk (*) indicate a p-value≤0.05; with two (**) when it is ≤0.01 and with three (***) when it is ≤0.001.</p

NR4A1 expression does not decrease after inhibition of p38, JNK and Src pathways.

<p>NIH3T3 cells were serum-starved overnight in 0.1% BSA and then pretreated for 1 h with DMSO or inhibitors SB203580 10 µM (A), SP600125 10 µM (B) and SU6656 2 µM (C), and then stimulated for indicated time periods with PDGF-BB (20 ng/ml); total cell lysates (TCL) were prepared and subjected to immunoblotting (Ib).</p

A small pool of the cytoplasmic NR4A1 translocates into the nucleus after long PDGF-BB stimulation.

<p>Cytoplasmic and nuclear fractions were prepared from NIH3T3 cells serum-starved overnight in 0.1% BSA and stimulated with PDGF-BB (20 ng/ml) for indicated periods of time and NR4A1 levels were analyzed by immunoblotting (Ib). The purity of the fraction were confirmed by immunoblotting for the nuclear marker Lamin and the cytoplasmic marker Tubulin.</p

PDGF-BB-induced NR4A1 expression is decreased after treatment with the NF-κB inhibitor BAY11-7082.

<p>NIH3T3 cells were serum-starved overnight in 0.1% BSA. Cells were pretreated for 4 h with DMSO or NF-κB inhibitor BAY11-7082 (10 µM) (A and B) and then stimulated for indicated time periods with PDGF-BB (20 ng/ml). mRNA was measured with quantitative RT-PCR (A) and total cell lysate (TCL) analyzed by immunoblotting (Ib) using NR4A1 antibody and tubulin, as loading control (B). Panel A shows the result of one out of three independent experiments performed; error bars indicate the standard deviation between three replicates. An asterisk (*) indicate a p-value≤0.05.</p

(A) cDNA microarray analysis of mRNA in Smad4-deficient MDA-MB-468 cells after infection with adenovirus expressing LacZ or Smad4 and stimulation with 2 ng/ml TGFβ1 or 300 ng/ml BMP7

Normalized mean expression values with error bars from triplicate microarray expressions are shown (arbitrary units). (B and C) Semiquantitative RT-PCR analysis of and mRNA in MDA-MB-468 cells as in A (B) or in response to 2 ng/ml TGFβ1 in HaCaT cells (C). Amplified cDNA sizes are in bp. (D) SIK and α-tubulin loading control protein profiles in response to 5 ng/ml TGFβ1 in HaCaT cells. Protein sizes are in kilodaltons. (E) Induction of nuclear and cytoplasmic levels of endogenous SIK by 5 ng/ml TGFβ1 in HaCaT cells (bar, 10 μm).<p><b>Copyright information:</b></p><p>Taken from "TGFβ induces SIK to negatively regulate type I receptor kinase signaling"</p><p></p><p>The Journal of Cell Biology 2008;182(4):655-662.</p><p>Published online 25 Aug 2008</p><p>PMCID:PMC2518705.</p><p></p