, 2001). C/EBP β, especially LAP1 and LAP2, can be phosphorylated at several sites by many different protein kinases, such as mitogen-activated GSK2126458 solubility dmso protein kinases, protein kinase A, protein kinase C, glycogen synthase kinase 3, and calcium/calmodulin-dependent protein kinases, with different effects on its transcriptional activity, depending on the phosphorylation site (Mahoney et al., 1992; Wegner et al., 1992; Trautwein et al., 1993, 1994; Piwien-Pilipuk et al., 2001, 2002). In particular, whereas phosphorylation

of rat C/EBP β by protein kinase A, protein kinase C or glycogen synthase kinase 3 on Ser240, which is located in the DNA-binding domain, has been reported to attenuate DNA binding and induce nuclear export, Ser105 phosphorylation of LAP isoforms is a key determinant of its transactivation capacity (Trautwein et al., 1993, 1994; Buck et al., 1999; Piwien-Pilipuk et al., 2001, GSK2118436 solubility dmso 2002). We therefore evaluated C/EBP β phosphorylation on Ser105 as a marker of transcriptional activity for this transcription factor. By using an antibody that specifically recognizes C/EBP β phosphorylated on Ser105, we observed that LAP1 is phosphorylated on Ser105 only in the nuclear compartment, implying

its transcriptional activation. From our co-immunoprecipitation experiments, we determined that LAP1 is essentially present in CGNs in its sumoylated form, both in the cytoplasm and in the nucleus, Idelalisib cell line and that the phosphorylated form is only nuclear and is only detected when neurons are kept in pro-survival conditions. The SUMOs serve as modifiers, exerting their effect by becoming conjugated to target proteins and stabilizing them (reviewed

by Lieberman, 2004). Sumoylation provides a rapid and efficient way to modulate the subcellular localization, activity and stability of a wide variety of protein substrates (Dorval & Fraser, 2007). C/EBPs, including C/EBP β, are well-known targets of SUMOs, which control their transcriptional activity by releasing, in rats, the inhibitory action of a conserved inhibitory domain that is a target for lysine sumoylation (Kim et al., 2002). Concerning C/EBP β isoforms, both LAP1 and LAP2 are potential targets of SUMO-2/3, but only LAP1 has been demonstrated to be conjugated to SUMO-2/3, as confirmed by our present results in CGNs. C/EBP β sumoylation has been shown to regulate its transcriptional activity, without influencing its subcellular localization (Eaton & Sealy, 2003). When CGNs were shifted to K5 medium to induce apoptosis, we observed a decrease in the LAP1 level and an increase in the LIP level in the nuclear compartment, and a decrease in the LAP2 level in the cytosolic fraction. Concomitantly, p-(Ser105)-LAP1 disappeared from the nuclear fraction.