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. 2011 Nov 17;6(11):e26674. doi: 10.1371/journal.pone.0026674

Figure 5. siRNA-mediated silencing of SOCS6 expression inhibits neurite-outgrowth.

Figure 5

(A) 3 µg of siRNA was transfected into PC12 cells and after 6 days of transfection, the cells were screened for SOCS6 knockdown by Western-blot analysis, using anti-SOCS6 antibody. Control-siRNA (green non-targeting) was used as a negative control. The same membrane was stripped and reprobed with anti-GAPDH antibody for protein loading control. (B) Morphology of SOCS6-siRNA transfected PC12 cells as compared to the control transfected cells after 6 days of transfection with NGF treatment under light microscope. (C and D) Neurite length was measured in randomly chosen cells (at least 8–10 different fields and approx 5 cells per field) transfected with either control or SOCS6 siRNA with or without IGF-1. Neurite lengths were measured by tracing individual neurites (as described in experimental procedures) and results are expressed as (C) average of total primary, secondary and tertiary neurite lengths or (D) average number of neurites per cell. Statistical significance of the difference was determined using ANOVA. The result shows the mean ± S.E. of n = 3 combined experiments (***p<0.001, **p<0.01, *p<0.05).