Fig. 2: Depletion of ABHD17B inactivates HSCs.

a Fraction of Bodipy positive (top) and total cells (bottom) after two days in the indicated conditions. DMSO and nortriptyline (Nor) are negative and positive controls for Bodipy, respectively. siRNAs targeting UBB are a control for transfection efficiency, and siRNAs targeting GAPDH are a negative control for Bodipy. Error bars represent mean ± SEM (n = 10 biological replicates). One-way ANOVA test. b mRNA expression quantified by qRT-PCR two days after HSCs were transfected with the indicated siRNAs. Data represent three independent experiments. Error bars represent mean ± SEM (n = 3 biological replicates). One-way ANOVA test. c ABHD17B expression quantified by qRT-PCR three days after HSC transfection. Data represent two independent experiments from two donors (5 and 6), which are further analyzed in (d–g). Each dot represents the mean of biological replicates for pooled non-targeing control (NTC siP; n = 4 biological replicates), single non-targeting control (NTC si5; n = 4), and other indicated groups (n = 2). ABHD17B siP are compared to NTC siP; ABHD17B si1 and ABHD17B si4 are compared to NTC si5. HSCs from donor 5 (d) and donor 6 (e) were transfected with the indicated siRNAs prior to serum starvation and stimulation with scar-in-a-jar conditions for 72 h. α−smooth muscle actin (αSMA, red) and collagen (orange) were visualized by immunofluorescence in the same field of view. Nuclei were stained with Hoechst (blue). Representative images from two independent experiments are shown. White bars indicate 50 µm. Quantification of αSMA fibers (f) and collagen area (g) from scar-in-a-jar assay. Data represent two independent experiments for each of the two donors (5 and 6). Each dot represents averaged result from n = 4 biological replicates. Data from donors 5 and 6 are combined for four independent experiments. Error bars represent mean ± SEM. One-way ANOVA test. h Members of the ABHD17 family were depleted using pooled siRNAs. COL1A1 levels were quantified by qRT-PCR three days after transfection. Error bars represent mean ± SEM (n = 8 biological replicates for control groups of donors 1 and 4; n = 7 for the control group of donor 3; n = 4 for all other groups). One-way ANOVA test. Source data are provided as a Source Data file.