Clone d3a7

Informed consents were obtained from all the subjects under approved institutional review board protocols at The University of Texas MD Anderson Cancer Center and at the University of Pittsburgh. Messenger RNA of target molecules in association with clinical outcome was studied in available data from the Profiling of Resistance patterns and Oncogenic Signaling Pathways in Evaluation of Cancers of the Thorax (PROSPECT) expression dataset^{62 (link)}. Tissue microarrays (TMAs) of 118 LUADs (65 female; 53 male) obtained from the University of Pittsburgh were stained for pSTAT3 and ERβ by immunohistochemistry. Using specific antibodies for pSTAT3 (Clone D3A7; 1:400 dilution; Cell Signaling Technology) and ERβ (MCA1974ST; 1:20 dilution; Abd Serotec), pSTAT3 was scored for the percentage of positive tumor cells and the intensity on a scale of 0–3. The H-score of 0–300 was calculated by multiplying the percentage score and the intensity score. ERβ was scored as the total score of both nuclear and cytoplasmic as described previously^{54 (link)}. Tumors were considered positive for pSTAT3 and ERβ if at least two cores per case were positive. Correlation between pSTAT3 and ERβ was statistically evaluated using Spearman correlation.

The study included 62 DLBCL samples in two tissue microarrays from 2000–2011. Twenty-five cases were from nodal sites and 37 from extranodal sites. Only tumors with enough tissues were included. Immunohistochemical stains of p-STAT3 (Y705, rabbit monoclonal Ab, clone D3A7, Cell Signaling) and p-SYK (Y525/526; rabbit polyclonal Ab; Cell Signaling) were performed and the results were evaluated by proportion of lymphoma cells that were stained. The antibody reactivity in ≥ 30% of the lymphoma cells was considered positive. Paraffin-embedded normal tonsil tissue was included as a baseline expression level for p-STAT3 and p-SYK.