Fig. 3: GBA1 expression, GCase protein levels, and lysosomal hydrolase activities in LN-229L444P cells following activation of TFs.
From: Direct and indirect regulation of β-glucocerebrosidase by the transcription factors USF2 and ONECUT2
a GBA1 and GBAP1 genes approximately to scale. Boxes: exons; lines: introns. Purple: 55-bp sequence unique to GBA1 exon 9. Pink: GBA1 RT-PCR primers. b Fold changes GBA1 mRNA (RT-qPCR) after CRISPRa of TFs in LN-229L444P cells. -actin (ActB) was used for normalization (N = 3–8 repeats). c GCase protein levels after CRISPRa of TFs in LN-229L444P cells. Cells were harvested 5 days post-transduction. d GCase protein quantification after CRIPSRa of TFs in LN-229L444P cells (N = 5–11 experiments). e Microscopy-based assessment of lysosomal hydrolase activities in live LN-229L444P cells. f GCase activity assessed by LysoFQ-GBA1 in live LN-229L444P cells after CRISPRa (N = 3–9 experiments). g α-N-acetylgalactosaminidase (NAGAL) activity in live LN-229L444P cells after CRISPRa of TFs. DGJNAc: NAGAL inhibitor (3 experiments). h Cathepsin B activity assessed in live LN-229L444P cells after CRISPRa of TFs. Bafilomycin A1: v-ATPase inhibitor (N = 3 experiments). i Hit selection process. Solid lines in C and D: medians, 25%, and 75% quartiles. Medians of NT controls: 100%. Unpaired t-test.
