Journal: Scientific Reports

Article Title: Anti-tumor activity of SL4 against breast cancer cells: induction of G 2 /M arrest through modulation of the MAPK-dependent p21 signaling pathway

doi: 10.1038/srep36486

Figure Lengend Snippet: MCF-7 and MDA-MB-231 cells were treated with SL4 as mentioned in . ( A ) CDK proteins, including Cyclin A2, Cyclin B1, phospho-Cyclin B1, cdc2 and phospho-cdc2(Tyr15), were detected by western blotting. β-actin expression was used as a loading control. ( B ) Protein expression levels of p-cdc2 and cdc2 (relative to β-actin) were determined. Mean ± SD for three replicate determinations. ( C ) CDK-regulated proteins, including p21, p53, Wee1, cdc25C and BRCA1, were detected by western blot. β-actin expression was used as a loading control. ( D ) Protein expression levels of cdc25C and p21 (relative to β-actin) were determined. Mean ± SD for three replicate determinations. ( E ) The mRNA expression levels of cdc25C and p21 were measured by quantitative PCR using specific primers as described in Materials and Methods. GAPDH was used as the control. Mean ± SD for three replicate determinations.

Article Snippet: The human breast cancer cell lines MCF7 (ER+, PR+, HER2-, p53 wild type, BRCA1 wild type), MDA-MB-231 (ER-, PR-, HER2-, p53 mutation, BRCA1 wild type), and MDA-MB-436 (ER-, PR-, BRCA1 mutation) were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Western Blot, Expressing, Control, Real-time Polymerase Chain Reaction

Journal: Carcinogenesis

Article Title: Insulin-like growth factor-I receptor is suppressed through transcriptional repression and mRNA destabilization by a novel energy restriction-mimetic agent.

doi: 10.1093/carcin/bgt251

Figure Lengend Snippet: Fig. 2. CG-5-induced suppression of IGF-IR expression is mediated through Sp1 downregulation and another β-TrCP-dependent mechanism. (A) Western blot analysis of the concentration-dependent effects of CG-5 on the expression of IGF-IR and various transcription factors related to IGF-IR gene regulation, including Sp1, WT1, p53 and BRCA1, in LNCaP cells in 10% FBS-supplemented RPMI 1640 medium (left) and the corresponding densitometric analyses of relative expression levels (right; n = 3). (B) Western blot analysis of the effect of siRNA-mediated knockdown of Sp1 on IGF-IR expression in LNCaP cells. (C) Concentration-dependent decreases in the recruitment of Sp1 to the IGF-IR gene promoter in response to CG-5 treatment in LNCaP cells as determined by ChIP analysis. IP, immunoprecipitation. (D) Ectopic expression of Sp1 protected against the suppressive effect of CG-5 on IGF-IR gene transactivation as determined by IGF-IR promoter–luciferase reporter assay (n = 3 independent experiments). Inset, immunoblot of Sp1 expression levels in LNCaP cells transiently transfected with a plasmid encoding Sp1 versus the pCMV control. (E) Western blot analysis of the concentration-dependent effect of CG-5 on Sp1 and IGF-IR expression in LNCaP cells transiently transfected with a plasmid encoding Sp1 versus the pCMV control (left) and the corresponding densitometric analysis of relative expression levels (right; n = 3). (F) Western blot analysis of the concentration-dependent effect of CG-5 on the expression of Sp1 and IGF-IR in LNCaP cells transiently transfected with a plasmid encoding Myc-tagged ΔF-β-TrCP, a dominant-negative form of β-TrCP, versus the pCMV control. All western blots shown are representative of three independent experiments.

Article Snippet: Antibodies against various proteins were obtained from the following sources: IGF-1R, Sp1, WT1, p53, BRCA1 and cyclin E from Santa Cruz Biotechnology (Santa Cruz, CA); Akt, p-473S-Akt, p-1135Y-IGF-1R, β-transducin repeat-containing protein (β-TrCP) and Myc from Cell Signaling Technology (Beverly, MA); FLAG from Sigma–Aldrich and β-actin from MP Biomedicals (Irvine, CA). mAbs against the N- and C-termini of HuR were obtained from Santa Cruz (#sc-5261; mouse mAb) and Cell Signaling (#12582; rabbit mAb), respectively.

Techniques: Expressing, Western Blot, Concentration Assay, Knockdown, Immunoprecipitation, Luciferase, Reporter Assay, Transfection, Plasmid Preparation, Control, Dominant Negative Mutation

Journal: BMC Gastroenterology

Article Title: A novel predictive model of microvascular invasion in hepatocellular carcinoma based on differential protein expression

doi: 10.1186/s12876-023-02729-z

Figure Lengend Snippet: Univariate logistic regression analysis of MVI occurrence in the training cohort

Article Snippet: Subsequently, the following primary antibodies were added: GPC3, CK19, vimentin, mutant p53, AFP, EGFR, VEGF (all sourced from Santa Cruz Biotechnology, USA), RRM1, and BRCA1 (both sourced from Abcam, UK), After incubation for 30 min, the cells were stained with ultraView Universal DAB Kit (Roche Diagnostics, GER).

Techniques:

Journal: BMC Gastroenterology

Article Title: A novel predictive model of microvascular invasion in hepatocellular carcinoma based on differential protein expression

doi: 10.1186/s12876-023-02729-z

Figure Lengend Snippet: Multivariate logistic regression analysis of MVI occurrence in the training cohort

Article Snippet: Subsequently, the following primary antibodies were added: GPC3, CK19, vimentin, mutant p53, AFP, EGFR, VEGF (all sourced from Santa Cruz Biotechnology, USA), RRM1, and BRCA1 (both sourced from Abcam, UK), After incubation for 30 min, the cells were stained with ultraView Universal DAB Kit (Roche Diagnostics, GER).

Techniques:

Journal: Stem cells (Dayton, Ohio)

Article Title: Spontaneous transformation of human adult nontumorigenic stem cells to cancer stem cells is driven by genomic instability in a human model of glioblastoma.

doi: 10.1634/stemcells.2006-0585

Figure Lengend Snippet: Figure 6. Aberrant expression of DNA re- pair and checkpoint activation proteins in HNGC-2 cells. HNGC-1 (A) and HNGC-2 (B) cells were stained for detection of DNA damage foci and spindle abnormalities using an array of phosphoantibodies to DNA repair and checkpoint proteins Chk1, Chk2, Brca-1, Brca-2, Mre11, Rad52, Smc1, and Ark-1 by immunofluorescence. The HNGC-2 cells were intensely positive for all the molecules except Brca2. The cells exhibited a high level of aberrant expression with Chk2 and Brca1. The panel shows diffused nuclear staining and no DNA damage foci in HNGC-1 cells; 63 objective. Abbreviation: HNGC, human neuroglial culture.

Article Snippet: The cells were incubated with the following primary antibodies for 2 hours at 4°C: CD133 (1:50; Miltenyi Biotec, Bergisch Gladbach, Germany, http://www. miltenyibiotec.com), nestin, musashi, bmi-1, Sox-2 (1:100; Cell Signaling Technology, Beverly, MA, http://www.cellsignal.com), nucleostemin (1:100; Chemicon, Temecula, CA, http://www. chemicon.com), p53 (1:100; Santa Cruz Biotechnology Inc., Santa Cruz, CA, http://www.scbt.com), phospho-p53, p21, Notch-1, Notch-2, Notch-3, Hes1, Hes2, Hes4 (1:100; Chemicon), ataxiatelangiectasia mutated (pATM), 53BP1, H2AX1, Chk1, Chk2, Brca1, Brca2, Mre11, RAD52, ARK-1 (1:20–1:100; Cell Signaling), and Smc1 (1:100; Calbiochem, San Diego, http://www.emdbiosciences.com).

Techniques: Expressing, Activation Assay, Staining, Immunofluorescence