Fig. 5: HuD, NRN1 and GAP43 knockdown in co-cultures containing FUSP525L MNs.
From: HuD impairs neuromuscular junctions and induces apoptosis in human iPSC and Drosophila ALS models
A, B Representative merged images of day 14 co-cultures obtained with FUSWT SkMCs and FUSP525L MNs transfected with the indicated siRNAs and stained with the indicated antibodies. Scale bar: 50 μm. Single panels are shown in Supplementary Fig. S7. C, D The graphs report quantitative analysis of the percentage of α-BTX positive fibers (C) and CC-3 fluorescence intensity (D), at day 14 of co-culture as in (A, B). Each dot represents an individual batch of differentiated iPSCs transfected with the indicated siRNAs. The following number of randomly selected fields were used for the α-BTX analysis: 5 (non-targeting siRNA replicate 1 and 2, HuD siRNA replicate 2); 7 (GAP43 siRNA replicate 3); 6 (all other replicates). The following number of randomly selected fields were used for the CC-3 analysis: 8 (non-targeting siRNA replicate 3, HuD siRNA replicate 1, NRN1 siRNA replicate 3); 9 (HuD siRNA replicate 2 and 3; non-targeting siRNA replicate 1); 10 (all other replicates). The dot shows the average value. Error bars indicate standard deviation calculated on the average value of the replicates. Ordinary one-way ANOVA, p = 0.0009 for α-BTX positive fibers, p = 0.0003 for CC-3 immunofluorescence; post hoc Tukey test p-values for multiple comparisons are indicated in the graph. Source data are provided as a Source Data file.
