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shRNA knockdown of Ankrd26, Trio, Gps2, HMMR and Dipa induces adipocyte differentiation in 3T3-L1 cells.

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posted on 20.02.2013, 04:30 by Xiu-Fen Liu, Tapan K. Bera, Charissa Kahue, Thelma Escobar, Zhaoliang Fei, Gregory A. Raciti, Ira Pastan

To down-regulate Ankrd26, Trio, Gps2, HMMR and Dipa expression cells were transfected with specific shRNA for Ankrd26 (Ankrd-sh), Trio (Trio-sh), Gps2 (Gps2-sh), HMMR (HMMR-sh), Dipa (Dipa-sh) or with non-targeting control shRNA (Co-sh). A) Ankrd26 mRNA expression determined by Real-Time Quantitative PCR analysis of total RNA isolated from 3T3-L1 cells transfected with Ankrd-sh or Co-sh. mRNA levels in Ankrd-sh treated cells are relative expression units to those in control (Co-sh; mean ± SD; n = 3). **p<0.01. Macroscopic images (B) and lipid quantification (C) in Ankrd-sh and Co-sh transfected cells stained with oil-red-O upon differentiation. Bars are expressed as means ± SEM of oil-red-O values measured at 490 nm. Ankrd-sh vs. Co-sh, **p<0.01. mRNA expression (D, G, J and M), macroscopic images (E, H, K and N) and lipid quantification (F, I, L and O) of 3T3-L1 cells transduced with Trio-sh (D–F), Gps2-sh (G–I), HMMR-sh (J–L) and Dipa-sh (M–O) were evaluated as A, B and C, respectively.