Figure 7

Detection of HLTF protein by immunofluorescence in 3 thyroid cancer cell lines and Immunodetection of HLTF on Western Blot. The 3 thyroid cancer cell lines derived from human thyroid cancers: papillary carcinoma B-CPAP (A), follicular carcinoma FTC-133 (B) and anaplastic carcinoma 8505C (C). HLTF was detected using the ART2 rabbit antiserum and revealed by a secondary antibody coupled to Alexa Fluor as described in the Methods. The scale bar is 50µm. (D-F) 40µg (E and F) or 60µg (D) of cell lysate (total [D], nuclear [E] and cytoplasmic [F]) were analyzed by 4%-12% gradient PAGE-SDS. After protein transfer to a membrane, immunodetection was performed with the α-HLTF rabbit serum (Sigma, 1/500 [E and F] and 1/1000 [D]) as described in Methods. Lane 1: TPC-1. Lane 2: B-CPAP. Lane 3: FTC-133. Lane 4: 8505C. Lane 5: HeLa.