Journal: Oncotarget

Article Title: Notch system is differentially expressed and activated in pituitary adenomas of distinct histotype, tumor cell lines and normal pituitaries

doi: 10.18632/oncotarget.19046

Figure Lengend Snippet: ( A – C ) Gene expression normalized to Gapdh in AtT20 cells and mouse pituitaries determined by qRT-PCR and expressed as percentage of change of the normal pituitaries which were considered 100%. *p = 0.0003 for Notch3, Notch1,2,4 NS, n = 3–5; 3–8 for AtT20 and pituitary respectively (A), p = 0.0111 and p = 0.0105 for the Notch ligands Dll1 and Jagged1 , respectively vs pituitary and (B) p = 0.0016, 0.0184 and 0.0032 for the Notch target genes Hes1, Hes5 and Hey2 , respectively, vs. pituitary n = 2–3; 3 for AtT20 and pituitary respectively (C). ( D , E ) Active and membrane domains of NOTCH1-3 receptor levels determined by Western Blot: Bars show the mean of the receptor normalized to β actin levels, expressed as the percentage of normal mouse pituitary. *p = 0.026 for NOTCH1, p < 0.0001 for NOTCH2 and p = 0.0146 for NOTCH3 vs. pituitary, for the active domains (80KDa) n = 3–6; 5–7 for AtT20 and pituitary respectively (D). No significant differences were found for the membrane domain (110 KDa) n = 3–6; 3–7 for AtT20 and pituitary respectively (E). Representative Western blots showing the active and membrane domains of NOTCH receptors in normal pituitaries (Pit) and ATt20 cells ( F ).

Article Snippet: For mouse tissue the following antibodies were used: Rabbit polyclonal antibody against NOTCH1 and NOTCH2 (dilution 1:700, Merck Millipore), NOTCH3 (dilution 1:200, Santa Cruz Biotechnology Inc), HES1 (dilution 1:400, Merck Millipore).

Techniques: Gene Expression, Quantitative RT-PCR, Membrane, Western Blot

Journal: Oncotarget

Article Title: Notch system is differentially expressed and activated in pituitary adenomas of distinct histotype, tumor cell lines and normal pituitaries

doi: 10.18632/oncotarget.19046

Figure Lengend Snippet: ( A – C ) Gene expression determined by qRT-PCR in pituitaries of lacDrd2KO and Drd2l oxP/loxP counterparts normalized to Gapdh and expressed as percentage of change of the Drd2 loxP/loxP pituitaries considered 100%. Bars show mean values. Notch1 *p = 0.0431, Notch3 *p = 0.0089 vs. Drd2 loxp/loxp ; Notch2, 4 NS, n = 7, 5 for lacDrd2KO and Drd2 loxP/loxP mice, respectively (A). Notch ligands Dll1 and Jagged1 NS (B), and Notch target genes Hes1 (* p = 0.0106 lacDrd2KO vs. Drd2 loxP/loxP ) , Hes5 and Hey2 , NS (C); n = 7, 5 for lacDrd2KO and Drd2 loxP/loxP mice, respectively. ( D – E ) The expression of the active and membrane domains of NOTCH1-3 receptors was determined by Western blot, related to the correspondent β actin levels, and expressed as the percentage of Drd2 loxP/loxP pituitary average. *p = 0.01 for NOTCH1, NOTCH2, 3 NS active domain, p =0.0604 and p = 0.0042 for NOTCH2 and NOTCH3 membrane domains, respectively for lacDrd2KO vs Drd2 loxP/loxP m ice. n = 4, 3–4 for lacDrd2KO and Drd2 loxP/loxP . Representative Western blots showing the active and membrane domains of NOTCH receptors in lacDrd2KO and Drd2 loxP/loxP ( F ). Representative microphotographies of NOTCH1-3 and HES1 Immunohistochemistry performed in lacDrd2KO and Drd2 loxP/loxP mice ( G ).

Article Snippet: For mouse tissue the following antibodies were used: Rabbit polyclonal antibody against NOTCH1 and NOTCH2 (dilution 1:700, Merck Millipore), NOTCH3 (dilution 1:200, Santa Cruz Biotechnology Inc), HES1 (dilution 1:400, Merck Millipore).

Techniques: Gene Expression, Quantitative RT-PCR, Expressing, Membrane, Western Blot, Immunohistochemistry