FIG. 4.
PTHrP/forskolin blocks the association of HDAC4 and 14-3-3 proteins and increases the association between HDAC4 and MEF2C. (A) USCs were transfected with a GAL4-firefly luciferase reporter and an SV40-renilla luciferase reporter plus expression vehicles encoding Gal4-HDAC4(2-740), 14-3-3-VP16, or Gal4-VP16 in either the absence or presence of PTHrP or forskolin as indicated. RLU, relative luciferase units. P, phosphate group. (B) USCs were transfected with a GAL4-firefly luciferase reporter and an SV40-renilla luciferase reporter plus expression vehicles encoding either Gal4-HDAC4(2-740) or MEF2C-VP16 in either the absence or presence of PTHrP or forskolin. (C) USCs were transfected with expression vehicles encoding HDAC4-Flag plus 14-3-3-HA in either the absence or presence of forskolin as indicated. 14-3-3-HA and associated proteins were immunoprecipitated, and Western analysis (WB) was employed to detect co-IP 14-3-3-HA and HDAC4-Flag. (D) USCs were transfected with expression vehicles encoding HDAC4-Flag plus MEF2C-HA in either the absence or presence of forskolin as indicated. HDAC4-Flag and associated proteins were immunoprecipitated and Western analysis was employed to detect co-IP HDAC4-Flag and MEF2C-HA.