Fig 2: Go and KEGG enrichment analysis of genes related to E2F2 in CRC tissues in the TCGA-COADREAD data sets. (A-C) Go enrichment analysis showed the BP (biological processes), CC (cellular components), and MF (molecular function) of co-expressed genes with E2F2. (D) Significantly enriched KEGG terms obtained from KEGG enrichment analysis of co-expressed genes with E2F2.

Fig 3: E2F2 expression level was reduced in clinical CRC tissues. (A-D) Representative images of E2F2 protein expression level in paraffin-embedded (A) normal mucosa and CRC tissues with (B) pathological stage I, (C) pathological stage II and (D) pathological stage III. Scales represent 100 micron. Representative images with original magnificent at 200×. (E) Quantifications of the average optical density (AOD) for E2F2 protein expression in normal mucosa and CRC tissues with pathological stage I-III. (F) Western blotting analysis and (G) quantitative real-time polymerase chain reaction analysis of five paired samples of CRC tissues and adjacent normal tissues from randomly selected CRC patients. * , p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Fig 4: Correlation of NELFE and E2F2 with the proportion of TICs. TIC profile in tumor samples and correlation analysis. (A) Barplot showing the proportion of 22 kinds of TICs in gastric cancer samples. The column names in the plot are sample ID. (B) Heatmap showing the correlation between 22 kinds of TICs and numeric cells in each small box indicating the p value of the correlation between two kinds of cells. The shade of each small color box represents the corresponding correlation value between two cells, and the Pearson coefficient was used for the significance test. (C) Violin plot showing the ratio differentiation of 22 kinds of immune cells between gastric cancer samples with low or high NELFE expression relative to the median NELFE expression level, and Wilcoxon rank sum was used for the significance test. (D) Venn plot displaying five kinds of TICs correlated with NELFE expression codetermined by difference and correlation tests displayed in violin and scatter plots, respectively. (E) Violin plot showing the ratio differentiation of 22 kinds of immune cells between gastric cancer samples with low or high E2F2 expression relative to the median E2F2 expression level, and Wilcoxon rank sum was used for the significance test. (F) Venn plot displaying six kinds of TICs correlated with E2F2 expression codetermined by difference and correlation tests displayed in violin and scatter plots, respectively. (G, H) Scatter plot showing the correlation of the proportions of 5 kinds of TICs with NELFE expression and the proportions of 6 kinds of TICs with E2F2 expression.

Fig 5: Lung protein content in O2/air recovered mice. In (A) lung E2F2, (B) cyclin A1, (C) p21, and in (D) phosphorylated eIF2α. Upper panels: densitometry values of WT (grey bars) and KO (open bars) normalized to Calnexin loading controls. Values are the mean±S.E. of 2. Lower panels: representative Western blots of each signal with calnexin as loading control. For eIF2α, the ratio of serine 51 phosphorylated signal over total protein content is shown. In E: level of serine 51 phosphorylated eIF2α over total in KO and WT MEF. Grey bars: WT, open bars: KO. Values are the mean±S.E. of 3 in each group. *: p<0.05 vs WT. Lower panel: representative Western blot of serine 51 positive signal over total. Calnexin is the loading control.