Figure 1
Lipidomic profiling of Rb-depleted MEFs. (a) RT–qPCR of Rb1 in wild-type MEFs infected with lentiviruses expressing the indicated shRNAs and selected. Cells were cultured in the medium containing 1% FBS for 24 h. Data represent mean+s.d. Four batches of MEFs derived from one embryo were used per one shRNA. **P<0.01 by Student’s t-test. (b) Schematic of strategy used to identify lipids altered in Rb-depleted MEFs. (c) PCA score plot of 199 lipids (n=4). (d) Hierarchical clustering of 199 lipids (n=4). (e) Top VIP scores and heat map from PLS-DA of 199 lipids. Red and green indicate increased and decreased levels, respectively. (f) Volcano plot of lipids with VIP>1. Important lipids were selected with fold change threshold (x) and p-values (t-test) threshold (y). Important variables with P<0.05 and |FC|>1.5 were indicated with red and those with P<0.01 and |FC|>2 were labeled. (g) Levels of the indicated lipids in control MEFs (solid) and Rb-depleted MEFs (blank). Data represent mean+s.d. (n=4) **P<0.01 by Student’s t-test. FBS, fetal bovine serum; PCA, principal component analysis; PLS-DA, partial least square discriminant analysis.
