Journal: Cell Stress & Chaperones

Article Title: Drug-induced keratin 9 interaction with Hsp70 in bladder cancer cells

doi: 10.1007/s12192-018-0913-2

Figure Lengend Snippet: Hsp70 and IgG antibodies were linked to Dynabeads, and T24 bladder cancer cell lysates were immunoprecipitated. SDS-PAGE, followed by silver staining, was performed and the band at the arrow was sliced from the gel and analyzed by LC-MS/MS. The unknown band was determined to be keratin 9. a Hsp70-Dynabead-linked Immunoprecipitation of T24 lysates in the absence of VER155008 (VER-) and presence of VER155008 (VER+). Lane C is an IgG-Dynabead-linked immunoprecipitate of a DMSO-treated T24 lysate (IgG control). Molecular weight markers (MWM) of 25, 50, and 75 represent molecular weights in kilodaltons. b bold black and red letters represent keratin 9 amino acid fragments identified from the excised band by LC-MS/MS. Red letters identify the amino acids found in the filament super family

Article Snippet: The protocol was followed according to the manufacturer’s instructions, beginning with the addition of 10 μL of Hsp70 antibody (StressMarq) or 10 μL of mouse IgG (Santa Cruz Biotechnology) to create the Co-IP bead complex.

Techniques: Immunoprecipitation, SDS Page, Silver Staining, Liquid Chromatography with Mass Spectroscopy, Molecular Weight

Journal: Cell Stress & Chaperones

Article Title: Drug-induced keratin 9 interaction with Hsp70 in bladder cancer cells

doi: 10.1007/s12192-018-0913-2

Figure Lengend Snippet: Western blot analysis of human bladder cancer cell lysates (T24) treated with DMSO, the Hsp90 inhibitor STA9090, the Hsp70 inhibitors VER155008, and MAL3-101 or combinations of the various inhibitors. The blot was probed with a keratin 9 mouse monoclonal antibody

Article Snippet: The protocol was followed according to the manufacturer’s instructions, beginning with the addition of 10 μL of Hsp70 antibody (StressMarq) or 10 μL of mouse IgG (Santa Cruz Biotechnology) to create the Co-IP bead complex.

Techniques: Western Blot

Journal: Cell Stress & Chaperones

Article Title: Drug-induced keratin 9 interaction with Hsp70 in bladder cancer cells

doi: 10.1007/s12192-018-0913-2

Figure Lengend Snippet: Immunostaining using keratin 9 and Hsp70 mouse monoclonal antibodies and visualized by confocal microscopy. T24 human bladder cancer cells were processed as described in “ .” The negative controls without primary antibodies ( a , keratin 9) and ( c , Hsp70) show slight background staining resulting from autofluorescence of the secondary antibody. The nuclei were stained with DAPI. Perinuclear distribution of keratin 9 ( b ) and punctate distribution of Hsp70 ( d )

Article Snippet: The protocol was followed according to the manufacturer’s instructions, beginning with the addition of 10 μL of Hsp70 antibody (StressMarq) or 10 μL of mouse IgG (Santa Cruz Biotechnology) to create the Co-IP bead complex.

Techniques: Immunostaining, Confocal Microscopy, Staining