Journal:

Article Title: Nrdp1-mediated regulation of ErbB3 expression by the androgen receptor in androgen-dependent but not castrate-resistant prostate cancer cells

doi: 10.1158/0008-5472.CAN-09-4440

Figure Lengend Snippet: (A, B) CWR22 xenograft tumors were established by subcutaneous injections of cell suspensions (2.5 × 106 cells in Matrigel; 1:1, v/v) bilaterally into the flanks of 4–5-week-old nu/nu athymic male mice (n=10) previously implanted with sustained-release testosterone pellets. When palpable tumors were observed, animals were treated with (i) vehicle (peanut oil) or (ii) the AR antagonist bicalutamide (n=5/group). After two weeks on this treatment, the mice were euthanized, tumors were harvested and divided into sections that were paraffin-embedded snap-frozen in liquid nitrogen. Paraffin-embedded tumors were analyzed by immunohistochemistry for ErbB3 whereas frozen tumors were excised, lysed and protein levels were determined by Western blotting. (A) Western blotting revealed that ErbB3 levels increased while Nrdp1 levels decreased with bicalutamide treatment. (B) Immunohistochemistry demonstrating that ErbB3 levels increased in tumors extracted from the bicalutamide-fed mice. Brown staining denotes ErbB3 while blue represents counterstaining with hematoxylin. Negative control showed no ErbB3 staining, the xenograft from the vehicle-fed mouse shown showed lower ErbB3 expression (scored +1) compared to the bicalutamide-treated one (scored +3). (C) In contrast, CWR22Rv1, a CRPC cell line derived from a CWR22 relapsed tumor grown in a castrated mouse, failed to respond to bicalutamide and in these cells, Nrdp1 and ErbB3 were not androgen regulated. (D) Scheme describing androgen regulation of Nrdp1 transcription in androgen-dependent but not androgen-independent cells.

Article Snippet: Suspensions of CWR22 cells were made in 50% Matrigel solubilized basement membrane (BD Biosciences, Bedford, MA) and xenografts were established by subcutaneous injections of 2.5 × 10 6 cells/site into both flanks.

Techniques: Immunohistochemistry, Western Blot, Staining, Negative Control, Expressing, Derivative Assay

Journal: PLOS Computational Biology

Article Title: Development of a scoring function for comparing simulated and experimental tumor spheroids

doi: 10.1371/journal.pcbi.1010471

Figure Lengend Snippet: a) 2D cross-sections of 3D multiphoton microscopy image stacks depicting one replicate of each collagen concentration . Spheroids were imaged at one, two or three days after embedding in collagen, and were then fixed and imaged (see Section 4.3). b) Deviation score comparison between all spheroid samples. For each day, the deviation scores for three replicates of each collagen concentration are shown on the upper triangle, and the average deviation score for each collagen concentration is shown on the lower triangle. A top-down view of the spheroid point cloud for a representative replicate of each collagen concentration is shown next to each heatmap (see for all replicates). Blue cells are classified as non-gaslike, and red cells are classified as gaslike. Due to matching initial conditions, we observe low deviation scores between spheroids grown for one day. These differences increase at day 2, where we observe an approximately linear increase of the average deviation score from c = 1 mg/ml to c = 4 mg/ml. Finally, at day 3, we observe the lowest deviation between c = 3 mg/ml and c = 4 mg/ml. This underlines the findings by Kang et al., who observed a transition in invasion behavior between 2 mg/ml and 3 mg/ml. c) Deviation score box plots from spheroids grown for one, two and three days respectively. The box plots for each day show the deviation score values between c = 1 mg/ml and each other concentration. These values correspond to those used for the lowest rows in b). We observe that for spheroids grown for one and two days, the deviation score values of c = 1 mg/ml compared with itself are similar to the deviation score values of c = 1 mg/ml compared with the other concentrations. The differences are nonsignificant and therefore not sufficient to clearly distinguish between the concentrations. This changes at day three, where each concentration shows a higher deviation score to c = 1 mg/ml than c = 1 mg/ml compared with itself. Significance was determined using Welch’s t-test.

Article Snippet: Once formed, individual spheroids were fully embedded into a 3D fibrous gel prepared using rat-tail collagen I (Corning, No. 354249) [ ].

Techniques: Microscopy, Concentration Assay, Comparison