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. 2015 Oct 26;125(12):4544–4558. doi: 10.1172/JCI82871

Figure 4. GPAT4, AGPAT3, and AGPAT5 contribute to SFA-induced mineralization and ER stress of VSMCs.

Figure 4

ShRNA-mediated knockdown of acyltransferases in VSMCs. MOVAS-1 cells were infected with lentiviruses containing acyltransferase shRNAs for 48 hours and selected with 5 μg/ml puromycin to generate each acyltransferase stable knockdown VSMC. Levels of acyltransferase mRNAs were determined by qPCR. Each shRNA reduced its targeted acyltransferase by over 75%. (A) Mineralization of VSMCs in the presence of 18:0. The stable acyltransferase knockdown VSMCs were treated with 200 μM 18:0 as BSA complex for 7 days in the presence of 2 mM inorganic phosphate. (B) Levels of Chop mRNA in acyltransferase stable knockdown VSMCs treated with 200 μM 18:0. The stable acyltransferase knockdown VSMCs were treated with 200 μM 18:0 as BSA complex for 24 hours. Levels of Chop mRNA were analyzed by qPCR. **P < 0.001 vs. MOVAS-1 cells treated with empty shRNA and vehicle. n = 4; #P < 0.01 and ##P < 0.001 vs. MOVAS-1 cells treated with empty shRNA and 200 μM 18:0 (one-way ANOVA).