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. 2017 May 8;8:15111. doi: 10.1038/ncomms15111

Figure 8. Genetic manipulation of TAp63α regulates lipid content in THLE2 hepatocytes.

Figure 8

(a) Representative dual channel fluorescent photomicrograph of THLE2 cells showing staining of lipids for oleic acid (BODIPY 493/503, green) and nuclei (DAPI, blue) of THLE2 cells after the over-expression of TAp63α or ΔNp63α. Magnifications 63X. Right panel shows total lipid content (green area) (n=4 per group). (b) Protein levels of TAp63α, ΔNp63α, XBP1s and FAS in THLE2 cells after the over-expression of TAp63α or ΔNp63α (n=4 per group). (c) ER fragments in HepG2 cells transfected with empty plasmid control and plasmid encoding TAp63α. (d) Representative dual channel fluorescent photomicrograph of THLE2 cells showing staining of lipids for oleic acid (BODIPY 493/503, green) and nuclei (DAPI, blue) of THLE2 cells cultured in oleic acid medium, treated with empty siRNA control (left image) or siRNA TAp63α isoform (right image). Magnifications 63X. Right panel shows total lipid content (green area). (e) Protein levels of TAp63α, pIKKβ, XBP1s and FAS in THLE2 cells exposed to oleic acid, after treatment with siRNA against either control or TAp63α (n=3 per group). (f) Representative dual channel fluorescent photomicrograph of THLE2 cells showing staining for oleic acid (BODIPY 493/503, green) and nuclei (DAPI, blue) of THLE2 cells transfected with empty plasmid control (left image), plasmid encoding TAp63α (middle image) and co-transfected with plasmid encoding TAp63α and siRNA IKKβ 24 h after transfection (right image). Magnifications 63X. Right panel shows total TG (green area). (g) Protein levels of TAp63α, pIKKβ, XBP1s and FAS in THLE2 cells after TAp63 over-expression followed by IKKβ silencing. GAPDH was used to normalize protein levels (n=3 per group). Uncropped blots of this Figure accompanied by the location of molecular weight markers are shown in Supplementary Fig. 18. Data are presented as mean±standard error mean (s.e.m.). Statistical differences are denoted by *P<0.05, **P<0.01 and ***P<0.001. For multiple comparison (a,b,f,g) a one way ANOVA followed by Bonferroni or Kruskal-Wallis test was performed. Student t-test was used in the other panels.