Figure 1.

Experimental workflow of miRNA mimic screening and follow-up. (1) The primary miRNA screen was performed in duplicate, assaying both GCase activity and cell viability in WT and N370S/N370S Gaucher fibroblasts. (2) miRNA candidates were chosen based on the GCase Z-score, with consideration of their effect on viability. (3) Results were confirmed by retesting selected miRNAs in both 384- and 96-well plates. Then, the top 5 miRNAs that upregulated and 3 miRNAs that down-regulated GCase activity were selected for further studies. (4) The mRNA relative expression of GBA1, SCARB2 and PSAP was evaluated by real-time PCR. (5) Changes in protein levels were investigated by Western blot. (6) Additional studies were performed on miRNA candidates identified in the previous step.