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. 2010 Dec 13;5(12):e15208. doi: 10.1371/journal.pone.0015208

Figure 4. Directional shifts in experimental outcome with inappropriate reference gene selection.

Figure 4

Total RNA was isolated over time (0–10 days) from density-arrested 3T3-L1 preadipocytes treated with MDI and assessed for reference and target gene expression by qRT-PCR during complete adipocyte differentiation. Fold changes in reference genes that fell within (A) and outside (B) the ΔCT ≤ +/−0.5 limits of suitability. Fold changes in PPARγ gene expression without (C) and with normalization to 18S, cyc, or TfR (D), 18S, cyc, or α-tubulin (E), and 18S, cyc, or GAPDH (F). Arrows indicate directional shifts relative to normalization to 18S.