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. 2012 May 14;287(26):22354–22366. doi: 10.1074/jbc.M112.376491

FIGURE 7.

FIGURE 7.

Arc mRNA is destabilized by translation-dependent RNA decay. A, quantitative analysis of Arc mRNA expression by real-time semiquantitative PCR shows Arc levels in cells stimulated with Cch for 3 h. Actinomycin D was applied in combination with wortmannin (wort) or vehicle (veh) 60 min after the start of continuous Cch treatment. Arc mRNA levels are normalized to both hypoxanthine-guanine phosphoribosyltransferase and cyclopholin and expressed as fold-change relative to vehicle treatment (p < 0.05; n = 6). B, representative Western blot and quantitative analysis show Arc protein expression 60 or 240 min after the start of continuous Cch treatment. Cells received wortmannin or vehicle 60 min after the start of Cch treatment. Arc protein levels are normalized to GAPDH and expressed in percent of Arc expression at 60 min post-Cch stimulation. Asterisks indicate a significant change between groups (n = 4, p < 0.05). C, representative Western blot shows Arc protein expression in cells stimulated with Cch for 240 min. Cells additionally received wortmannin and/or anisomycin (ANI) 1 h after onset of stimulation. Additional controls are provided in Fig. S5.