A) WB analyses with antibodies specified on the right of total cell extracts from HEL cells untreated or exposed for 24 h either to U0126 [100 μM] or PD98059 [100 μM], as indicated. GAPDH was used as loading control. Exposure to U0126, but not to PD98059 [100 μM], reduced the levels of ERK phosphorylation and the content of HDAC5 and GATA1 but did not affect the content of HDAC4 and HDAC1.
B) WB analyses with antibodies specified on the right of total cell extracts from K562 cells untreated or exposed for 24 h to either U0126 [100 μM] or PD98059 [100 μM] (Exp I) and U0126 [100 μM] or Wortmannin [100 μM] (Exp II), as indicated. GAPDH was used as loading control. Exposure to U0126, but not to PD98059 or wortmannin, reduced the levels of ERK phosphorylation and HDAC5, GATA1 and EKLF content without affecting HDAC1 content.
C) WB analyses with antibodies specified on the right of nuclear extracts from K562 cells untreated or exposed to U0126 [100 μM] for 1, 2, 4, 6 and 24 h, as indicated. Laminin B was used as loading control. Exposure to U0126 reduced the nuclear levels of ERK phosphorylation within 1 h and the nuclear content of HDAC5, GATA1 and EKLF by 6 h. HDAC5 was still detected but GATA1 and EKLF became barely detectable after 24 h of exposure to the inhibitor.