Validation of rabbit monoclonal antibody D60C5 by Western blotting.

<p>Positions of molecular weight standards (kilodaltons) are shown to the left of each blot. A, HeLa cells harboring Tet-On construct inducible with doxycycline. B, Comprehensive uterine cancer cell line panel (endometrial and cervical). Note: C4I harbors biallelic mutations of LKB1: a chromosomal deletion plus a point mutation that does not affect protein levels <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073449#pone.0073449-Wingo1" target="_blank">[1]</a>. CaSki, C33, and ME180 do not harbor LKB1 mutations <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073449#pone.0073449-Wingo1" target="_blank">[1]</a>. Endo was derived from normal endocervical epithelium immortalized with HPV E6/E7 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0073449#pone.0073449-Fichorova1" target="_blank">[27]</a>.</p

Validation of LKB1 rabbit monoclonal antibody D60C5 for immunohistochemistry in human and mouse paraffin-embedded, formalin-fixed samples.

<p>A, Human cervical cancer cell lines. HeLa/Lkb1  =  HeLa cells following transduction of lentivirus harboring a human <i>LKB1</i> cDNA inducible expression construct. Relative expression levels of Lkb1 protein are indicated in parentheses. B, Mouse tissues (endometrium and lung) from animals harboring floxed alleles of <i>Lkb1</i> following Cre-mediated recombination with <i>Sprr2f-Cre</i> (endometrium) or nasal-instillation of Adeno-Cre virus (lung). Distinct Lkb1-null clones are indicated by dashed lines (endometrium) or arrows (lung). Bars = 10 µm for each panel. Asterisks in the lung panels show invasive cancer cells subjacent to the dysplastic epithelium; these invasive cancer cells are also clearly Lkb1-null. C, Percent of Lkb1-null cells in <i>Sprr2f-Cre</i>; <i>Lkb1^L</i>^/<i>L</i> female mice by immunohistochemistry at 3, 6, 12, and 20 weeks of age. Error bar  =  S.E.M. D, Normal patterns of LKB1 protein in human lung and oviduct highlighting localization to the apical surface of ciliated cells (arrows). Note: in the oviduct, ciliated epithelial cells (arrows) are interspersed among nonciliated cells. Bars = 10 µm in both panels.</p

Summary of patient data.

<p>Summary of patient data.</p

Testing of another α-LKB1 monoclonal antibody (Ley 37D/G6).

<p>Tissue sections are from the uterus of a 6-week old <i>Sprr2f-Cre</i>; <i>Lkb1^L</i>^/<i>L</i> female mouse. Rabbit monoclonal D60C5 readily distinguishes LKB1 positive from negative cells as shown previously. In contrast, the mouse monoclonal antibody Ley 37D/G6 shows a homogeneous pattern throughout the endometrial epithelium (serial step section) and fails to distinguish between LKB1 positive and negative cells. Size bars = 100 µ for each panel.</p

Scoring schema for LKB1 and pAMPKα (Thr172) expression in human lung cancer specimens.

<p>Tissues were paraffin-embedded and fixed in formalin. Only staining in the malignant epithelial cells was scored. A, LKB1 immunohistochemistry and representative cases illustrating histologic scores. B, pAMPKα (Thr172) immunohistochemistry and representative cases illustrating histologic scores. The dynamic range was somewhat lower for pAMPKα (Thr172) vs. LKB1 but a wide range of staining intensities was also observed. Bar = 10 µm in all panels; all panels are at same magnification.</p

PDAC tumor engraftment in mice is associated with poor patient outcomes.

<p>(A) Kaplan-Meier curves of recurrence-free survival and tumor engraftment for patients with resectable primary tumors and no evidence of metastatic disease. (B) Kaplan-Meier curves of overall survival and tumor engraftment for the entire study population. (C) Kaplan-Meier curves of recurrence-free survival and rate of tumor engraftment for patients with resectable primary tumors and no evidence of metastatic disease. (D) Kaplan-Meier curves of overall survival and rate of tumor engraftment for the entire study population.</p

Pathological characteristics of 133 patients with resected pancreatic ductal adenocarcinoma by tumor engraftment status.

<p>Pathological characteristics of 133 patients with resected pancreatic ductal adenocarcinoma by tumor engraftment status.</p

Baseline demographic and clinical characteristics of 57 patients with patient-derived PDAC xenografts based on xenograft growth rate.

<p>Baseline demographic and clinical characteristics of 57 patients with patient-derived PDAC xenografts based on xenograft growth rate.</p

Recurrence-free and overall survival by tumor engraftment and xenograft growht rate.

<p>Recurrence-free and overall survival by tumor engraftment and xenograft growht rate.</p

Molecular characteristics of patient tumors do not predict tumor engraftment.

<p>(A) Genetic analysis of <i>KRAS</i> and <i>TP53</i> in patient tumors. Amino acid changes resulting from mutations are listed. Samples for which mutations were not detected are indicated by a dash. (B) Representative positive and negative immunohistochemical staining of SMAD4 in primary patient tumors. Staining of stromal cells on each slide served as a positive control. Scale bars = 100 μm. (C) Summary of SMAD4 expression in patient tumors that were successfully or unsuccessfully engrafted in mice.</p