Journal: Cancer cell

Article Title: Loss of p53 in enterocytes generates an inflammatory microenvironment enabling invasion and lymph node metastasis of carcinogen-induced colorectal tumors.

doi: 10.1016/j.ccr.2012.11.014

Figure Lengend Snippet: Figure 4. Activation of NF-kB Is Associated with Cxcl1 and Twist Upregulation in p53-Deficient Invasive Carcinoma (A) GSEA comparing all differentially regulated genes from AOM-induced invasive cancers in Tp53DIEC mice with a data set of NF-kB target genes obtained from microdissected invading human tumor epithelia (Horst et al., 2009). Normalized enrichment score: 1.46; p < 0.0001. (B) Immunoblot analysis for the indicated proteins in lysates from invasive cancers from Tp53DIEC mice and noninvasive adenomas from Tp53F/F mice 16 weeks after the first AOM administration. (C–F) Immunohistochemical analysis of activated p-p65 (C), CXCL1 (D), F4/80 (E), and Gr-1 (F) in invasive cancers of AOM-challenged Tp53DIEC mice. Dashed white lines mark the basal membrane. Scale bars represent 50 mm. (G) Immunoblot analysis for the indicated proteins in lysates from invasive cancers from Tp53DIEC mice and noninvasive adenomas from Tp53F/F mice 16 weeks after the first AOM administration. (H) Immunohistochemical analysis of Twist in invasive cancers of AOM-challenged Tp53DIEC mice. The scale bar represents 50 mm. (I) Colocalization of E-cadherin (green) and vimentin (red) in invasive cancer epithelial cells of AOM-challenged Tp53DIEC mice, indicating EMT; nuclei are stained using 40,6-diamidino-2-phenylindole (blue). The scale bar represents 200 mm. (J–M) Representative immunohistochemical analysis of p53 (J and L) and p-p65 (K and M) in human colon cancer samples. Scale bars represent 100 mm. (N) Correlation of p53 staining and p-p65 staining (n = 59; c2 test, p < 0.05). (O) Correlation of p-p65 staining and the presence of lymph node (LN) metastases (n = 59; c2 test, p < 0.02). (P) Relative CD68 mRNA level in human colon cancer patients who were characterized by expression of both p53 and p-p65 (p53+/p-p65+; n = 23) or not (other; n = 30). Data are mean ± SE; *p < 0.05 by t test. (Q and R) Representative immunohistochemical staining of CD68 in human colon cancer patients who were characterized by expression of both p53 and phospho-p65 (Q) or the absence of p53 and phospho-p65 (R). Scale bars represent 100 mm. See also Figure S3.

Article Snippet: Standard immunohistochemical procedures were performed using the following antibodies: b-catenin (Santa Cruz; sc-1496), bromodeoxyuridine (BrdU) (AbD Serotec; MCA 2060), cleaved caspase-3 (Cell Signaling; 9661), CXCL1 (R&DSystems;MAB453), F4/80 (Caltag; MF48000), Gr-1 (eBioscience; 12-5931-82), phospho-Histone H2A.X (Cell Signaling; 2577), E-cadherin (Becton Dickinson; 610182), Twist (Abcam; 49254), vimentin (Santa Cruz; SC-7557), VE-cadherin (Santa Cruz; SC-6458), and occludin (Invitrogen; 711500).

Techniques: Activation Assay, Western Blot, Immunohistochemical staining, Membrane, Staining, Expressing

Journal: British journal of pharmacology

Article Title: A novel compound, denosomin, ameliorates spinal cord injury via axonal growth associated with astrocyte-secreted vimentin.

doi: 10.1111/j.1476-5381.2012.02211.x

Figure Lengend Snippet: Figure 7 Denosomin-mediated axonal outgrowth in primary cultures is mediated by vimentin secretion from astrocytes. (A–C) Isolated astrocytes were cultured in the presence or absence of 1 mM denosomin (Pre-Deno) for 6 days. Rat spinal cord cells were cocultured on the astrocyte layer without denosomin for 7 days after the astrocytes had been treated with denosomin. (D–F) Isolated astrocytes were cultured in the presence or absence of 1 mM denosomin (Pre-Deno) for 6 days. The culture medium was then replaced with fresh medium without denosomin, and the medium was collected 24 h later for use as astrocyte-conditioned medium (ACM). Rat primary cultured spinal cord cells were cultured in normal medium or ACM for 6 days (F). (G and H) Cultured astrocytes that were treated with or without denosomin (Deno) (G) and ACM collected from the astrocyte culture (H) were used for the ELISA assay. (I–K) Isolated mouse cortical cells were cultured with or without 1 or 10 ng·mL-1 vimentin for 6 days. The cells were cultured on slides with (J and K) or without (I–J) a CSPG coating. The cells were immunostained for pNF-H and MAP2, and the densities of pNF-H-positive axons on each neuron were quantified (B, D, I and K). Cellular distributions of neurons and non-neuronal cells per 0.1 mm2 were evaluated by the quantification of MAP2- and DAPI-positive cells (E). In (B, G and H): *P < 0.05, Student’s unpaired t-test (two-tailed); #P < 0.05; one-way ANOVA followed by the post hoc Bonferroni test, versus the control ACM treatment (D), versus the control cells (I) or versus CSPG-coated control cells (K). The numbers of photos (B, D, E, I, and K) or measurements (G and H) are shown in parentheses in the columns. The scale bar indicates 100 mm.

Article Snippet: The vimentin concentrations in these samples were quantified using a rat vimentin ELISA kit according to the manufacturer’s protocol (Cusabio Biotech, Wuhan, Hubei, China).

Techniques: Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Control

Journal: PLoS ONE

Article Title: Isolation of putative stem cells present in human adult olfactory mucosa

doi: 10.1371/journal.pone.0181151

Figure Lengend Snippet: Primary antibodies.

Article Snippet: Anti-Vimentin , Bioss , bs-0756R , Rabbit , 1:300 , .

Techniques: