Patatin-Like Phospholipase Domain-Containing Protein 3 Abundance is Affected by Knockdown of Sterol Regulatory Element-Binding Protein 1c but not of Carbohydrate Response Element Binding Protein.

Kendall, Sophia J.; White, Heather M.

Patatin-like phospholipase domaincontaining protein 3 (PNPLA3) is a key lipase involved in fatty liver syndrome onset and recovery in humans and bovine. Regulation of PNPLA3 is mediated by carbohydrate response element binding protein (ChREBP) in rodents and sterol regulatory element-binding protein 1c (SREBP1c) in humans. The objectives of our study were to optimize an siRNA knockdown of ChREBP and SREBP1c and to determine if either transcription factor knockdown would affect PNPLA3 abundance in an immortalized bovine hepatocyte cell line, BFH12. Hepatocytes were cultured in monolayer and at least 80% confluent after 24h when cells were randomly assigned in triplicate to treatment with ChREBP-targeted (siC1, siC2, siC3) or SREBP1c-targeted (siS1, siS2, siS3) siRNA at either 20, 60, or 80μM, or a nonspecificsiRNA control (siNON). Media was replenished after 4h and cells incubated for 20h before harvest in TRIzol for subsequent protein isolation. Protein abundances were determined by Western blot and normalized to total lane protein. Data were analyzed (PROC MIXED, SAS 9.4) with fixed effect of siRNA and concentration. Differences were declared at P≤0.05; tendencies at 0.05.

STEROL regulatory element-binding proteins; CARRIER proteins; CARBOHYDRATES; PHOSPHOLIPASES; PROTEINS; TRANSCRIPTION factors

Journal of Animal Science, 2022, Vol 100, p270

0021-8812

Academic Journal

10.1093/jas/skac247.490