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. 1998 Jun 15;18(12):4451–4460. doi: 10.1523/JNEUROSCI.18-12-04451.1998

Fig. 7.

Fig. 7.

Fibrillar Aβ stimulates rapid, transient phosphorylation of CREB on serine 133 in THP1 monocytes and microglia that is blocked by piceatannol and PD98059. A, THP1 monocytes were stimulated with 50 μm Aβ25–35 for the indicated times. B, Primary cultures of rat microglia were stimulated with 50 μm Aβ25–35 for 2 min.C, Aβ-induced phosphorylation of CREB is blocked by pretreatment of THP1 monocytes with piceatannol. THP1 monocytes were pretreated with 50 μg/ml piceatannol or an equal amount of DMSO (final concentration, 70 mm) for 1 hr, followed by stimulation with 50 μm Aβ25–35 for 2 min.D, Aβ-induced phosphorylation of CREB is blocked by pretreatment of THP1 monocytes with PD98059. THP1 monocytes were pretreated with 25 μm PD98059 or an equal amount of DMSO for 20 min, followed by stimulation with 50 μm Aβ25–35 for 2 min. Phosphorylation of CREB in A–D was determined by Western blot analysis on RIPA lysates (100 μg of protein) using anti-phospho-CREB polyclonal antibodies (top panel). The blots were stripped and reprobed using anti-CREB antibody (bottom panel).E, Pretreatment of THP1 monocytes with PD98059 does not inhibit tyrosine phosphorylation. THP1 monocytes were pretreated with 25 μm PD98059 or an equal amount of DMSO for 20 min, followed by stimulation with 50 μm Aβ25–35 for 2 min. Western blot analysis of tyrosine phosphoproteins was performed on RIPA lysates (100 μg of protein) using monoclonal anti-phosphotyrosine antibodies (4G10).