Blood. 2006 March 1; 107(5): 2170–2179.

Figure 3. Overexpression of Ezh2 in HSCs. (A) WBC counts of mice given transplants with BM cells transduced with control () or Ezh2 () vectors (n = 11 recipients per group, 2 independent experiments). Mean values plus or minus standard error of the mean (SEM) are shown. (B) Chimerism at different time points after transplantation. The percentage of donor-derived transduced CD45.1+GFP+ WBCs is shown. Average values ± 1 SEM of 2 independent experiments are shown. (C) Ezh2 protein expression in the spleen about 120 days after primary transplantation of control or Ezh2 CD45.1 BM cells. (D) CAFC frequencies of sorted LSK GFP+ cells 120 days after primary transplantation (control, ; Ezh2, ). (E) Chimerism levels of secondary recipients, competitively transplanted with various ratios (2:1, ⋄; 1:1, ○; 1:2, □; 1:5, ▵) of transduced/nontransduced and freshly isolated BM cells, analyzed 3 months after transplantation. Values show data from individual recipients in 2 independent experiments (n = 35/group). (F) CRI calculated for CD45.1+GFP+ (transduced) versus CD45.1+GFP- (nontransduced) cells (see insert). Values (+ 1 SEM) are averages 3 months after secondary transplantation, based on 11 and 24 individual mice in the first and second experiment, respectively (control, ; Ezh2, ). (G) CRI calculated for transduced cells (CD45.1+GFP+) compared to freshly isolated BM cells (CD45.2+; see insert). Averages values (+ 1 SEM) of 11 and 24 individual mice of the first and second experiment, respectively, are shown (control, ; Ezh2, ). (H) LTRA frequencies in CD45.1+GFP+ (transduced) cell fractions calculated from limiting dilution analyses 3 months after secondary transplantation in 2 independent experiments (control, ; Ezh2, ).

Am J Pathol. 2008 July; 173(1): 30–41.

Figure 4 HGF blocks IL-1β- or TNF-α-mediated RANTES expression in tubular epithelial cells. A: HGF blocked RANTES expression induced by IL-1β and TNF-α. Human proximal tubular epithelial cells (HKC-8) were treated with IL-1β (5 ng/ml), TNF-α (2 ng/ml), HGF (40 ng/ml), or in different combinations as indicated for 48 hours. Whole cell lysates were immunoblotted with antibodies against RANTES and actin, respectively. B: HGF inhibited IL-1β-induced RANTES expression in a dose-dependent manner. HKC-8 cells were incubated with IL-1β (5 ng/ml) alone, or with IL-1β plus increasing amounts of HGF as indicated. C: HGF inhibited IL-1β-mediated RANTES secretion. The supernatant of the HKC-8 cells after various treatments as indicated was immunoblotted with anti-RANTES antibody. D: HGF inhibited TNF-α-induced RANTES secretion. The levels of RANTES secreted in the supernatant of HKC-8 cells after different treatments were measured by a specific enzyme-linked immunosorbent assay.

Am J Pathol. 2008 July; 173(1): 68–76.

Figure 6 Phenotypic score of WT mice injected with G-CSF or vehicle compared to KO mice. Mice were treated as indicated and both flanks were irradiated with 30 Gy. At the indicated times after irradiation, mice were evaluated for a skin reaction according to the phenotypic scale given in Materials and Methods. A and B represent two independent experiments. Values are the mean ± SEM from five mice per group scoring two flanks per mouse. *P < 0.05 for WT (V) versus WT (GCSF); +P < 0.05 for KO versus WT (V). Am J Pathol. 2008 July; 173(1): 68–76.

Am J Pathol. 2008 July; 173(1): 68–76.

Figure 1 Influx of neutrophils and macrophages into flank skin of Smad3 WT and KO mice at various times after irradiation. After irradiation flank skin was processed at long times (up to 160 days) (A, B) or short times (up to 48 hours) (C, D) and neutrophils (A, C) and macrophages (B, D) were identified by immunostaining as described in Materials and Methods. The numbers of stained cells in five ×400 fields were counted for each irradiated flank and results are expressed as mean ± SEM. The numbers of flanks processed at each time point were four per genotype in A and B and eight per genotype in C and D. *P < 0.05 for WT (white bars) and KO (black bars) at the same time point. E: Immunostaining of neutrophils in WT (left) and KO (right) dermis 8 hours after irradiation. Arrows point to clusters of neutrophils stained with rat anti-mouse neutrophil antibody and visualized with brown DAB chromagen. Notice the large numbers of neutrophils surrounded by arrows and located in the bottom right corner of the KO (right) panel. DAB with hematoxylin. Original magnifications, ×400. Am J Pathol. 2008 July; 173(1): 68–76.

Reproductive Biology and Endocrinology 2008, 6:41

Quantitative RT-PCR of KLF9 mRNA in human endometrium and endometrial tumors. A normalized cDNA panel of human endometrial tumors was obtained from OriGene Technologies, Inc. This panel was comprised of cDNAs from n = 6 of normal (N) endometria, n = 9 of Stage I tumors, n = 8 of Stage II tumors, n = 19 of Stage III tumors, and n = 6 of Stage IV tumors. Shown are box plots (median, upper and lower quartiles, minimum and maximum data values) of relative abundance of KLF9 mRNA for tumors (delineated by stage and tumor type: endometrioid, serous). Sample numbers are indicated. ANOVA indicated no differences in mRNA abundance between any of the individual stages. However, a significant difference between combined stages (normal plus stage I vs. stages II, III, and IV) was noted by the Mann-Whitney Rank Sum test. Simmen et al. Reproductive Biology and Endocrinology 2008 6:41

Reproductive Biology and Endocrinology 2008, 6:36

Immunostaining results of CD45, CD68 and PAF-R. CD45 (top), CD68 (middle) and PAF-R (bottom) immunostaining results in stroma, as assessed by image analysis (top) and manual scoring (middle, bottom) in cervical samples from controls (C) (n = 18), responders (R) (n = 13) and non-responders (NR) (n = 9). The "box-and-whisker plot" represents the median value with 50% of all data falling within the box. The whiskers extend to the 5th and 95th percentiles. Boxes with different letter designations are significantly different, p < 0.05. Sahlin et al. Reproductive Biology and Endocrinology 2008 6:36

J. Clin. Invest. 118(1): 40-50 (2007)

Adrenomedullin signaling is necessary for murine lymphatic vascular developmentJ. Clin. Invest. Kimberly L. Fritz-Six, et al. 118:40 doi:10.1172/JCI33302 [Go to this article.] Figure 8AM signaling preferentially mediates enhanced ERK activation in HMVEC-dLys compared with HUVECs. (A) Cultured HMVEC-dLys and HUVECs are morphologically and genetically distinct cell lines based on histology and expression pattern of Prox1. Original magnification, ×400. (B) Stimulation of HUVECs and HMVEC-dLys with the potent growth factor VEFGA resulted in a dose-dependant increase in cell proliferation that was not significantly different between the 2 cell lines. Data represent averages of 4 independent experiments, each performed in duplicate. (C) Stimulation of HUVECs and HMVEC-dLys with AM peptide resulted in a dose-dependant increase in cell proliferation that was significantly greater in HMVEC-dLys compared with HUVECs. *P < 0.05. Data represent averages of 4 independent experiments, each performed in duplicate. (D) Stimulation of HUVECs and HMVEC-dLys with 10 nM AM peptide resulted in a significantly greater induction of ERK phosphorylation in HMVEC-dLys compared with HUVECs over a 30-minute time course. *P < 0.03 at 15- and 20-minute time points. Data represent averages of 3 independent experiments, each performed in duplicate. (E) Induction of ERK activation by AM stimulation in HMVEC-dLys was significantly reduced by the RAMP2-specific peptide inhibitor AM22-52 and completely blocked by the MAPK inhibitor PD98057 (PD). *P < 0.05 compared with untreated; #P < 0.05 compared with AM-treated. Data represent averages of 3 independent experiments, each performed in duplicate.

J. Clin. Invest. 118(1): 64-78 (2007)

The receptor tyrosine kinase EphA2 promotes mammary adenocarcinoma tumorigenesis and metastatic progression in mice by amplifying ErbB2 signalingJ. Clin. Invest. Dana M. Brantley-Sieders, et al. 118:64 doi:10.1172/JCI33154 [Go to this article.] Figure 3Elevated EphA2 expression in MCF10A. ER2 cells enhances cell proliferation and invasiveness in vitro. (A) Parental MCF10A human breast cells and MCF10A.HER2 cells were transduced with adenoviruses (Ad) expressing EphA2 or control β-gal and plated on growth factor–reduced Matrigel to generate 3-dimensional spheroid cultures. After 10 days in culture, parental MCF10A cells and cells expressing Ad–β-gal formed small, round acinar structures, while MCF10A.HER2 cells formed larger colonies with irregular, invasive protrusions (arrows). Expression of Ad-EphA2 in MCF10A cells resulted in larger, irregular colonies, an effect that was amplified in MCF10A.HER2 cells (P < 0.05; single-factor ANOVA). Scale bar: 25 μm. (B) Cultures were stained with TO-PRO-3 iodide nuclear stain (red) and anti-Ki67 (green) and imaged by confocal microscopy. Confocal analysis revealed that parental and Ad–β-gal–transduced MCF10A formed uniform acinar structures composed of a single layer of epithelial cells surrounding a central lumen, while MCF10A.HER2 cells formed multiacinar structures with invasive protrusions (arrows) and a poorly defined lumen containing several cells. MCF10A cells transduced with Ad-EphA2 also formed multiacinar structures with a poorly defined lumen. Invasion and lumen filling were enhanced in MCF10A.HER2 cells overexpressing EphA2. Scale bar: 20 μm. EphA2 overexpression significantly enhanced proliferation (Ki67+ nuclei, arrows) within acinar structures formed by MCF10A and MCF10A.HER2 cells (P < 0.05; single-factor ANOVA). (C) Expression of adenoviral gene products and overexpression of ErbB2/HER2 in MCF10A.HER2 cells was confirmed by immunoblot, and uniform loading was verified by immunoblot for actin. Expression of p-Erk, total Erk, p-EphA2, and total EphA2 was also assessed by immunoblot.

Arthritis Res Ther. 2008; 10(3): R53

Figure 5 Gastrointestinal AE discontinuations with different therapies in three analyses of trials lasting 4 to 12 weeks. AE, adverse event; NSAID, nonsteroidal anti-inflammatory drug. Arthritis Res Ther. 2008; 10(3): R53.

J Clin Invest. 2008 January 2; 118(1): 100–110

Figure 4 Detachment sensitizes skin carcinoma cells specifically to TRAIL and CD95L. (A) Subconfluent cells were incubated in the absence (control) or presence of TRAIL (iz-muTRAIL, 1 μg/ml) and/or bortezomib (20 nM) for 24 hours. Cell death was determined by PI staining and flow cytometric analysis. (B and C) Cells were trypsinized and cultured either on an uncoated (attached) or a poly-HEMA–coated surface (detached) in the absence or presence of TRAIL (B; 1 μg/ml) or at the indicated concentrations (C). (D) Subconfluent cells (DT02) were treated with or without 1 μM thapsigargin (Tgg), 30 μg/ml oxaliplatin (Oxp), 5 μg/ml cycloheximide (Chx), or 43°C heat shock (HS) or irradiated with 15 Gy (IR) and were incubated in the absence or presence of TRAIL (1 μg/ml) for 24 hours. (E) Attached (att) or detached (det) cells were incubated in the absence or presence of TRAIL (1 μg/ml) or CD95L (100 ng/ml) for 24 hours. Each data point represents one of the cell lines DT01–DT06 or DT11. The mean percentage of dead WT cells is indicated by a horizontal line. (F) Attached or detached cells were incubated in the absence or presence of TRAIL (1 μg/ml), CD95L (100 ng/ml), 5-FU (200 μg/ml), or etoposide (10 μM) for 24 hours. Standard deviation for duplicates or triplicates is shown. Results are representative of at least 2 independent experiments. J Clin Invest. 2008 January 2; 118(1): 100–110.