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. 2018 Apr 17;131(8):jcs211482. doi: 10.1242/jcs.211482

Fig. 7.

Fig. 7.

Reduction of endogenous SMN causes a reduction in cytoplasmic NCDN foci in SH-SY5Y cells. (A) SH-SY5Y cells were transfected with plasmids to express shRNAs targeting SMN (shSMN), Cyclophilin B (shCyclophilin) or with the empty pSuper GFP vector (data not shown), fixed after 72 h, and immunostained for endogenous NCDN and SMN allowing detection of NCDN foci within the cytoplasm (chevron arrowheads), as well as SMN-positive nuclear gems (triangular arrowheads). Images are single deconvolved z-sections. Scales bars: 7 µm. (B) The depletion of SMN results in a reduction in the number of NCDN foci present in the cytoplasm to 15.3 (±7.2) (mean±s.d.) from 20.6 (±12.0) and 19.5 (±7.6) compared to cells transfected with either shCyclophilin B or the empty pSuper GFP vector, respectively (ANOVA, P<0.0005, n=64 from 3 replicates). The box represents the 25–75th percentiles, and the median is indicated. The whiskers show the range of the data excluding outliers identified by the Tukey method. Outliers are shown as individual points. (C) The depletion of SMN causes a reduction of nuclear gems to 0.17 (±0.38) from 2.3 (±0.84) or 3.0 (±1.0) compared to cells transfected with shCyclophilin B or empty vector respectively confirming efficient reduction in SMN protein levels. Results are mean±s.d. (n=30 from three replicates). *P≤0.05, **P≤0.01, ***P≤0.001, ****P≤0.0001 (ANOVA).