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PAX6, TP53, TP63, FOSL2, FOXC1, PPARγ and NFκB mRNA expression (A) and PAX6 and FOSL2 protein levels (B) in LECs transfected with FABP5 siRNA or control (ctrl) siRNA. Western blot image from a single biological replicate (C) . Data are shown as mean ± SEM. Statistical analysis has been performed using two-way ANOVA followed by Tukey test (n = 7). p-values below 0.05 were considered statistically significant. PAX6 mRNA level was significantly lower in FABP5 knockdown LECs than in ctrl siRNA LECs, in KSFM medium or following LPS and IL-1β treatment (p ≤ 0.022). Furthermore, PAX6 protein level was significantly lower following FABP5 knockdown, than in ctrl siRNA LECs without treatment and after IL-1β treatment (p ≤ 0.031). TP63 mRNA level was significantly higher in ctrl siRNA LECs following LPS or IL-1β treatment, than without treatment (p ≤ 0.015). FOSL2 mRNA level was significantly higher in ctrl siRNA LECs following IL-1β treatment, than without treatment (p = 0.033). Under LPS and IL-1β treatment, FOSL2 mRNA levels were significantly lower in FABP5 knockdown cells than in control (transfected) cells (p ≤ 0.025).

Journal: PLOS One

Article Title: Fatty acid-binding protein 5 (FABP5) modulates limbal epithelial cell homeostasis by regulating the expression of key genes under both normal and inflammatory conditions , in vitro

doi: 10.1371/journal.pone.0347228

Figure Lengend Snippet: PAX6, TP53, TP63, FOSL2, FOXC1, PPARγ and NFκB mRNA expression (A) and PAX6 and FOSL2 protein levels (B) in LECs transfected with FABP5 siRNA or control (ctrl) siRNA. Western blot image from a single biological replicate (C) . Data are shown as mean ± SEM. Statistical analysis has been performed using two-way ANOVA followed by Tukey test (n = 7). p-values below 0.05 were considered statistically significant. PAX6 mRNA level was significantly lower in FABP5 knockdown LECs than in ctrl siRNA LECs, in KSFM medium or following LPS and IL-1β treatment (p ≤ 0.022). Furthermore, PAX6 protein level was significantly lower following FABP5 knockdown, than in ctrl siRNA LECs without treatment and after IL-1β treatment (p ≤ 0.031). TP63 mRNA level was significantly higher in ctrl siRNA LECs following LPS or IL-1β treatment, than without treatment (p ≤ 0.015). FOSL2 mRNA level was significantly higher in ctrl siRNA LECs following IL-1β treatment, than without treatment (p = 0.033). Under LPS and IL-1β treatment, FOSL2 mRNA levels were significantly lower in FABP5 knockdown cells than in control (transfected) cells (p ≤ 0.025).

Article Snippet: After centrifugation at 200g for 5 minutes, cells were cultured in Keratinocyte growth medium (KSFM) (Cat. Nr. C-20111, Promocell, Heidelberg, Germany), at 37 oC, 5% CO 2 .

Techniques: Expressing, Transfection, Control, Western Blot, Knockdown