Figure 3. Knockout of Lin37 results in a de-repression of cell cycle genes in G₀/G₁.

The NIH3T3 parental cell line as well as independent wild-type (WT, n = 4) and Lin37 knockout (KO, n = 6) single cell clones were arrested in G₀ by serum deprivation and re-stimulated for 9, 18, and 27 hr. (A) Cell cycle distribution was measured by PI staining and flow cytometry. One representative example of wild-type (Lin37^+/+) and knockout (Lin37^−/−) cells is shown. (B) mRNA of G₁/S (yellow) and G₂/M (blue) cell cycle genes was measured by qPCR in all cell lines with two technical replicates each and normalized to U6 expression. All data points for each gene were further normalized to the maximum mRNA expression of the parental cell line which was set to 100%. Mean values ± SD are given, and significances were calculated by the Students T-Test (*p≤0.05, **p≤0.01, ***p≤0.001).