Journal: Clinical Cancer Research

Article Title: FOXA1 Is a Potential Oncogene in Anaplastic Thyroid Carcinoma

doi: 10.1158/1078-0432.ccr-08-3155

Figure Lengend Snippet: Fig. 1. FOXA1 expression in human thyroid carcinoma cells. FOXA1 is expressed in the nuclei (N) but not in the cytoplasm (C) of human undifferentiated thyroid carcinoma 8505c cells as confirmed by Western blot analysis (A) and immunofluorescence staining detected through Texas red-conjugated secondary antibody (magnification, ×20; D). B, bright-field image of 8505c cell monolayer. C, nuclear staining by 4′,6-diamidino-2-phenylindole. MCF-7 (E) and MDA-MB-231 (F) human breast cancer cells represented the positive and negative controls for FOXA1 immunostaining, respectively. G, FOXA1 relative mRNA levels were significantly decreased in 8505c cells infected with sh#1 (average of the relative mRNA levels, 0.05 ± 0.03; ***, P < 0.001) and sh#2 (average of the relative mRNA levels, 0.66 ± 0.05; **, P < 0.01) versus control. H, 8505c cells were infected at the confluence of 60% for 3 to 6 h with two specific shRNAs anti-FOXA1 (sh#1 and sh#2) or shRNA anti-GFP sequence (sh-control). Nuclear FOXA1 protein levels were completely depleted with sh#1 and decreased by ∼30% with sh#2. Mean ± SD of at least three independent experiments.

Article Snippet: Materials and Methods Antibodies The following antibodies were used: β-actin (Sigma), lamin A/C (Cell Signaling Technology), FOXA1 (Ab5089 and Ab23738) for Western blot (Abcam), FOXA1 clone 2F83 for immunohistochemistry and Western blot (Seven Hills Bioreagents), p27kip1 (Transduction Laboratories), cleaved poly(ADP-ribose) polymerase (Cell Signaling Technology), cyclin D1 (Thermo Scientific), MIB-1 (DAKO), p21 (BD Pharmingen), and cyclin E1 (Santa Cruz Biotechnology).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Immunostaining, Infection, Control, shRNA, Sequencing

Journal: Clinical Cancer Research

Article Title: FOXA1 Is a Potential Oncogene in Anaplastic Thyroid Carcinoma

doi: 10.1158/1078-0432.ccr-08-3155

Figure Lengend Snippet: Fig. 2. FOXA1 expression in human primary thyroid undifferentiated carcinomas cells (ATC) and FISH analysis (all magnification, ×40). H&E stains of ATC cases showing squamoid morphology (A), classic giant cell and spindle cell morphology (D). B and E, intense nuclear FOXA1 expression in ATC cells by immunohistochemistry. Note the high mitotic activity (arrows). FOXA1 copy number gain by polysomy was observed in interphase tumor nuclei of ATC specimens (C and F) expressing high protein levels by immunohistochemistry (B and E). RP11-606C5 BAC clone (red) was cohybridized with a control RP11-314P15 BAC clone (green). C and F, representative sections from ATC with copy number gains of FOXA1. Intense FOXA1 nuclear staining is detected by immunohistochemical analysis (B and E).

Article Snippet: Materials and Methods Antibodies The following antibodies were used: β-actin (Sigma), lamin A/C (Cell Signaling Technology), FOXA1 (Ab5089 and Ab23738) for Western blot (Abcam), FOXA1 clone 2F83 for immunohistochemistry and Western blot (Seven Hills Bioreagents), p27kip1 (Transduction Laboratories), cleaved poly(ADP-ribose) polymerase (Cell Signaling Technology), cyclin D1 (Thermo Scientific), MIB-1 (DAKO), p21 (BD Pharmingen), and cyclin E1 (Santa Cruz Biotechnology).

Techniques: Expressing, Immunohistochemistry, Activity Assay, Control, Staining, Immunohistochemical staining