Article Figures & Data
Figures
- Figure 1.
Polyinosinic-polycytidylic acid (PolyIC) induced retinoic acid-inducible gene-I (RIG-I) expression on cultured human podocytes. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) (A, B) and western blot (C, D) analysis showed an increased expression of RIG-I after polyIC treatment. 500 ng/ml polyIC up-regulated mRNA of RIG-I (A) and 500 μg/ml PolyIC induced protein of RIG-I (C) in time dependent manner. Similarly, polyIC induced mRNA (B) and protein (D) of RIG-I in dose dependent manner. GAPDH was used as an internal control. The results represent the means±SEM (n=3 wells). *p<0.05; **p<0.001 versus control.
- Figure 2.
PolyIC induced melanoma differentiation-associated gene 5 (MDA5) expression on cultured human podocytes. qRT-PCR (A, B) and western blot (C, D) analysis showed an increased expression of MDA5 after polyIC treatment. Five hundred ng/ml polyIC up-regulated mRNA of MDA5 (A) and 500 μg/ml PolyIC induced protein of MDA5 (C) in a time-dependent manner. Similarly, polyIC induced mRNA (B) and protein (D) of MDA5 in dose dependent manner. GAPDH was used as an internal control. The results represent the means±SEM (n=3 wells). *p<0.05; **p<0.001 versus control.
- Figure 3.
polyIC increased the gene expression of inflammatory cytokines. Cultured human podocytes were treated with 500 ng/ml polyIC for 6 h. qRT-PCR analysis showed that the gene expression levels of interferon -β (IFN-β) (A) and interleukin-6 (IL-6) (B) were increased by polyIC treatment. GAPDH was used as an internal control. The results represent the means±SEM (n=3 wells). *p<0.05; **p<0.001 versus control.
- Figure 4.
Knockdown of RIG-I and MDA5 with siRNA inhibited the expression of inflammatory cytokines by polyIC. Cultured human podocytes were transfected with non-targeting (control) siRNA, RIG-I, and MDA5 siRNA using Dharmafect Reagent®. PolyIC (500 ng/ml) was added and incubated for 6 h. qRT-PCR analysis showed the gene expression levels of IFN-β (A) and IL-6 (B). GAPDH was used as an internal control. The results represent the means±SEM (n=3 wells). *p<0.05; **p<0.001.
- Figure 5.
PolyIC-induced actin reorganization was partially attenuated by RIG-I and MDA5 knockdown. F-actin representative microscopic images in human podocytes. Podocytes were transfected with non-targeting siRNA (cont si), RIG-I siRNA, or MDA5 siRNA, followed by treatment with 500 ng/ml poly IC for 24 h. At 72 h post-transfection, podocytes were fixed and stained with phalloidin to detect F-actin. Magnification 400×. Bar represents 50 μm.
