Figure 6

ILF3 regulates redox dependent mRNA stabilization in rMC-1 Müller cells. A) rMC-1 cells were transfected with siRNAs directed against Ilf3, Elavl1 or Hnrnpd, as indicated. Scrambled siRNA was used as control (Ctrl). Medium was changed 48 hours after transfection and then cells were (SD) or were not (C) serum deprived. H₂O₂ (50 μM) was added to the indicated samples. RNA levels were determined by real-time PCR two hours after treatment and expressed relative to untreated cells (C), which were set to 1 for each knockdown series. N = 4 to 8. B) Top panel: rMC-1 cells were transfected with siRNAs directed against Ilf3, Elavl1 or Hnrnpd, as indicated. Scrambled siRNA was used as control (Ctrl). Medium was changed 48 hours after transfection and respective target RNA levels were determined after two hours by real-time PCR. Bottom panels show protein levels at 48 hours after siRNA transfection. N = 4. C) rMC-1 cells were transfected with siRNAs directed against Ilf3, Elavl1 or Hnrnpd, as indicated. Scrambled siRNA was used as control (Ctrl). Medium was changed 48 hours after transfection and Lif RNA levels were determined after two hours by real-time PCR. To knockdown Ilf3 expression, two different siRNAs (Ilf3 (si1), Ilf3 (si2)) were used to exclude off-target effects. Shown are means ± SEM of N = 4. One-way ANOVA with Dunnett’s posttests was used to compare Lif levels after SD + H₂O₂ treatments (A) and in untreated knockdowns against control (C). Student’s t-test was used to compare downregulation of target genes (B). (**) P <0.01, (***) P <0.001. ANOVA, analysis of variance; ILF3, interleukin enhancer binding factor 3; SEM, standard error of the mean.