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Biomax MR films (Sigma) were scanned, and the estimated intensity of the bands was quantified using the Optiquant Bumetanide software package (Packard, Meriden, CT). A cohort of 38 patients with locally advanced rectal carcinoma received preoperative radiochemotherapy before surgery using a total dose of 45 Gy in daily fractions of 1.8 Gy on 5 consecutive days per week. During the first and fifth weeks of radiotherapy, 5-fluorouracil was delivered at a dose of 350 mg/m2 per day; and folinic acid was delivered at a dose of 20 mg/m2. The TNM was assessed by ultrasonography before radiotherapy, and the ypTNM staging was determined based on the surgical specimens. (ypTNM describes the anatomic extent of disease after surgery.) Two groups of patients (patients who were responsive and nonresponsive to radiotherapy) were determined by comparing the ultrasonographic TNM with the ypTNM. After treatment, the tumor stage grouping (ypTNM) in the responsive group was changed in all patients [19 (100%) of 19] because no more tumoral tissue was observed (complete regression). In the nonresponsive group, of 19 patients, the ypT stage was decreased for 7 (36.8%), unchanged for 10 (52.6%), and increased for the remaining 2 (10.5%). Moreover, Alisertib cell line in the 19 patients, the ypN stage was decreased in 9 (47.4%), unchanged in 6 (31.6%), and increased in 4 (21.1%). The ethics committee of the medical faculty and the state medical board agreed to these investigations and an informed consent was obtained from all patients. Tissues were immediately fixed for 24 hours in Bouin's fluid and handled as previously described.25 The Envision+ kit (Dako, Trappes, France) was used with anti-TP53 (1:800; BD Biosciences, Le Pont de Claix, France), anti-DIABLO (1:2000; Santa Cruz Biotechnology, Santa Cruz, CA), and anti-cleaved CASP8 (1:150; Ozyme, Saint Quentin Yvelines, France) antibodies. The CSAII (Biotin-Free Tyramide Signal Amplication) system (Dako) was used with anti-cleaved CASP9 antibodies (1:400; Clinisciences, Montrouge, France), and an LSAB kit (Dako) was used with anti-XIAP (1:80; BD Biosciences) and anti-cleaved CASP3 (1:80; Ozyme) antibodies. Two independent pathologists (M.D. and D.M.), Cobimetinib blinded to the clinicopathologic variables, evaluated the immunostaining. The immunostaining score was evaluated based on the whole slide. The results were recorded by assessing the percentage of positive cells and the intensity of the staining (1, mild; 2, moderate; 3, intense) on the tumoral and the nontumoral areas for each slide, as previously described.26 Immunostaining scores were calculated by multiplying the percentage of labeled cells by the intensity of staining. In vitro experiments were realized in quadruplicate and were performed at least three times in independent experiments. A one-way analysis of variance for dependent groups was performed to determine whether there were differences between all groups, followed by a Bonferroni post hoc test, if P