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. 2018 Nov 22;19(12):3716. doi: 10.3390/ijms19123716

Figure 4.

Figure 4

PKA activation following SA is dependent on soluble adenylyl cyclase (sAC). (a) Primary neurons at 15 DIV treated with Bic/4-AP (50µM/2.5mM, 10 min) with or without 20 min pre-treatment with the adenylyl cyclase (AC) inhibitors KH7 (20 µM), SQ22536 (SQ, 20 µM), and NKY80 (NKY, 20 µM) were subjected to immunoblotting with anti- phospho-PKA substrate (pPKA sub), phospho-CREB (pCREB), total CREB, and actin antibodies. (Ctrl was without pre-treatment) (b) Quantification of WB as in (a) showing the ratios pPKA sub/actin (c) and pCREB/CREB (d) expressed as a percentage of control (n = 4). (c) Primary neurons at 15 DIV treated with Bic/4-AP (2 h) with or without 20 min pre-treatment with the AC inhibitors KH7 (20 µM), SQ (20 µM), and NKY (20 µM) were subjected to immunoblotting with anti- Arc, c-Fos, EgrI, and actin antibodies. Results are representative of at least four experiments. Results show mean ± SD. One-way ANOVA followed by Bonferroni’s post-hoc test were used for evaluation of statistical significance. * p < 0.05, ** p < 0.01, *** p < 0.001.