Figures & data
Figure 1. (A) Recovery of DNA-dependent protein kinase (DNA-PK) activity after heat treatment of hybrid cells at 44 °C for 15 min with (■) or without (●) cycloheximide. Non-treated cell DNA-PK activity was set to 100%. Data are the average of two independent experiments. Error bars show the range. (B) Typical Western blot result of HSP72 and β-actin after heat treatment of hybrid cells with and without cycloheximide (CHX).
Figure 2. (A) Western blots of Ku80 bound HSP72, HSC73 and Ku80. (B) Ratios of Ku80 bound HSC73 (●) and HSP72 (■) following co-immunoprecipitation of nuclear extracts from hybrid cells with a Ku80 antibody. Hybrid cells were treated at 44 °C for 15 min, then cultured at 37 °C for the indicated time periods. Band densities were measured from photographs by a densitometer. Data are the average of two independent experiments. Error bars show the range.
Figure 3. Recovered DNA-dependent protein kinase (DNA-PK) activity (●) and Ku80 bound HSC73 concentrations (■) in nuclei from hybrid cells. Cells were treated at 44 °C for 15 min, then cultured at 37 °C for the indicated time periods.
Figure 4. (A) Western blot showing increased HSC73 and β-actin expression in hybrid cells carrying pcDNA3.1-HSC73. Lane 1: control hybrid cells; lane 2: hybrid cells carrying the pcDNA3.1 vector; lane 3: hybrid cells carrying pcDNA3.1-HSC73. (B) Recovery of DNA-dependent protein kinase (DNA-PK) activities during incubation at 37 °C after heat treatment at 44 °C for 15 min. Hybrid cells were transfected with pcDNA3.1-HSP73 (●) or the pcDNA3.1vector (■). Non-treated hybrid cell DNA-PK activity was set to 100%. Data are the average of two independent experiments. Error bars show the range.
Figure 5. (A) Western blots showing reduced HSP expression in hybrid cells carrying pSilencer-HSC73 or pSilencer-HSP72. Anti-HSC73 and β-actin: lane l, control hybrid cells; lane 2, hybrid cells transfected with pSilencer-HSP72; lane 3, hybrid cells transfected with pSilencer-HSC73. Anti-HSP72 and β-actin: control hybrid cells, or cells transfected with pSilencer-HSP72 or pSilencer-HSC73 for 2–10 h at 37 °C after heat treatment at 44 °C for 15 min. C is control cells. (B) Recovery of heat-inactivated DNA-dependent protein kinase (DNA-PK) activity in hybrid cells transfected with pSilencer-HSC73 (■) or pSilencer-HSP72 (●). Non-treated hybrid cell DNA-PK activity was set to 100%. Data are the average of two independent experiments. Error bars show the range.
