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Ning (no staining for p-ERK (score 0); weak p-ERK (score 1+), moderate p-ERK (score 2+), and strong p-ERK (score 3 +) staining) in Fig. 3. Staining intensities were grouped as dichotomous variables, defining scores 0? as low and 2? as high expression levels [25]. Evaluation of staining was carried out independently by two pathologists (MM and MA) blinded to patients' outcome and pathological stage
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Ning (no staining for p-ERK (score 0); weak p-ERK (score 1+), moderate p-ERK (score 2+), and strong p-ERK (score 3 +) staining) in Fig. 3. Staining intensities were grouped as dichotomous variables, defining scores 0? as low and 2? as high expression levels [25]. Evaluation of staining was carried out independently by two pathologists (MM and MA) blinded to patients' outcome and pathological stage
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A, c ?200; b, d ?Quadri et al. BMC Cancer (2017) 17:Page 5 ofFig. 3 Photomicrographs of pancreatic cancer tissue microarray (TMA) cores of p-ERK immunohistochemical staining: a no staining for p-ERK (score 0); b weak p-ERK (score 1+) staining; c moderate p-ERK (score 2+) staining; d strong p-ERK (score 3+) staining. Magnification: a-d: ?70 cases as 2+ (58.3 ), and 16 cases as 3+ (13.3 ) (Fig. 5a).
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AdeQuadri et al. BMC Cancer (2017) 17:Page 6 ofFig. 5 a Comparison of CNKSR1 expression of study cohort and secondary validation cohort. b Cellular distribution pattern of CNKSR1 showed primarily cytoplasmic expression in pancreatic cancer specimens. Nuclear staining of CNKSR1 was not associated with cytoplasmic CNKSR1 expression levels (0, 1+ vs 2+, 3+; p = 0.22; chi square test, 2-tailed)tumors
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Y intensity of p-ERK immunostaining. Figure 8C shows nuclear p-ERK expression levels (0, 1+, 2+, 3+)) by CNKSR1 cellular distribution (cytoplasmic CNKSR1 expression only vs cytoplasmic and nuclear) in pancreas cancer specimens of the SEER Pancreatic Cancer TMA (Mann Whitney U test; p = 0.017). To test whether expression levels of the two proteins are correlated as well, including if a possible neg
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Rbance values of the other wells. Average corrected absorbance was compared between transfectant and parental cells, using a t-test.ECM attachment assaysThe U87MG human glioma cell line was kept in tissue culture in DMEM (Cellgro Mediatech, Inc.), with 10 fetal bovine serum, and penicillin/streptomycin. For transfection, 2.5x106 cells were plated overnight on a 100 mm round dish. Cells were trans
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Thanized and their brains removed and either frozen using Optimal Cutting Temperature (OCT) compound (Electron Microscopy Sciences, Fort Washington, PA) in cryomolds placed atop dry ice, or fixed in 10 buffered formalin and paraffin embedded for histopathological analysis. The symptom-free survival of nude mice harboring U87MG parental versus transfected xenografts was compared. Kaplan-Meyer surv