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. 2019 Jan;95(1):1–10. doi: 10.1124/mol.118.112854

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Copyright © 2018 by The American Society for Pharmacology and Experimental Therapeutics

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Fig. 4. — Effect of CP55940, LDK1285, LDK1288, or LDK1305 on ERK1/2 phosphorylation over time. HEK293 cells expressing CB₁ were untreated (DMSO only) or treated with (A) CP55,940 (0.5 μM), (B) LDK1285 (10 μM), (C) LDK1288 (10 μM), or (D) LDK1305 (10 μM), for 2, 5, 10, 15, or 20 minutes. (E) Cells expressing CB₁ and either β-arrestin 1 siRNA or control siRNA were treated with DMSO vehicle only, LDK1285, LDK1288, or LDK1305 (all at 10 μM) for 5 minutes. Cell lysates were separated by SDS-PAGE and analyzed by Western blots probed for phospho-ERK1/2 (pERK1/2). The total level of ERK1/2 was detected for comparison. Note that the two bands correspond to the predominant isoforms of ERK, ERK1 (p44) and ERK2 (p42). Cells not expressing CB₁ and treated with LDK1288 showed no kinase phosphorylation relative to vehicle alone treatment (data not shown). Quantification of at least three independent experiments is displayed in the right-hand column and expressed as mean S.E.-fold increase above the basal level of phosphorylation.

Inline graphic — Effect of CP55940, LDK1285, LDK1288, or LDK1305 on ERK1/2 phosphorylation over time. HEK293 cells expressing CB₁ were untreated (DMSO only) or treated with (A) CP55,940 (0.5 μM), (B) LDK1285 (10 μM), (C) LDK1288 (10 μM), or (D) LDK1305 (10 μM), for 2, 5, 10, 15, or 20 minutes. (E) Cells expressing CB₁ and either β-arrestin 1 siRNA or control siRNA were treated with DMSO vehicle only, LDK1285, LDK1288, or LDK1305 (all at 10 μM) for 5 minutes. Cell lysates were separated by SDS-PAGE and analyzed by Western blots probed for phospho-ERK1/2 (pERK1/2). The total level of ERK1/2 was detected for comparison. Note that the two bands correspond to the predominant isoforms of ERK, ERK1 (p44) and ERK2 (p42). Cells not expressing CB₁ and treated with LDK1288 showed no kinase phosphorylation relative to vehicle alone treatment (data not shown). Quantification of at least three independent experiments is displayed in the right-hand column and expressed as mean S.E.-fold increase above the basal level of phosphorylation.