Fig. 1
SNCA-4x mDA neurons have a distinct but modulable morphological profile. (A) SNCA-4x and SNCA-corr mDA neurons were differentiated until D33, treated with 5 µM Prostratin or DMSO for 72 h and immunofluorescently stained. Image channels were segmented for morphological feature quantification. (B) Schematic overview of image data generation and morphological feature types. (C) Raw morphological feature values in control, and untreated and Prostratin treated SNCA-4x mDA neurons. Each datapoint represents technical replicate data from one 384-well plate well. Larger markers represent the medians each independent experiments. The boxplots show the median and the 2nd and 3rd quartiles of the data. Whiskers indicate 1.5X of the interquartile range. Welch’s unequal variances t-test was performed using the medians of independent differentiation experiments. ns = not significant, * = p < 0.05, ** = p < 0.01, *** = p < 0.001. (D) Scaled well and replicate median feature values aggregated into morphological profiles. Cosine similarity was used for clustering. (E) Top ten absolute mean differences between features of SNCA-4x and Control or SNCA-4x neurons treated with Prostratin. Means with standard deviations are shown. (F) Two first principal components (PCs) explaining most variation in the dataset after Principal Component Analysis (PCA). Each data point represents mean data from one well. (G) Absolute Pearson correlation of all features with the first ten PCs after PCA. Representative images illustrate cellular αSyn staining patterns. All data in this figure was generated in four independent experiments each containing at least 12 technical replicates per condition.
