Fig. 4: ABHD17B interacts with pathways involving contractility, adhesion, ECM, and the cytoskeleton. | Nature Communications

Fig. 4: ABHD17B interacts with pathways involving contractility, adhesion, ECM, and the cytoskeleton.

From: Screening the human druggable genome identifies ABHD17B as an anti-fibrotic target in hepatic stellate cells

Fig. 4: ABHD17B interacts with pathways involving contractility, adhesion, ECM, and the cytoskeleton.The alternative text for this image may have been generated using AI.

a RNA-seq and differential expression analysis were performed using nontargeting control (NTC) si5, and two siRNAs targeting ABHD17B (Supplementary Data 7). Dot plot displays the Gene Ontology (GO) terms most enriched in the genes repressed with depletion of ABHD17B. The color of each dot represents the adjusted p-value (hypergeometric test with Benjamini-Hochberg multiple testing correction), and the size of the dot represents gene count. b Heatmaps show expression of the repressed gene set for indicated GO categories for HSCs transfected with NTC (C), siRNA4 targeting ABHD17B (s4) and siRNA1 targeting ABHD17B (s1). Expression is centered and scaled by row (gene). c Proteins that interact with ABHD17B were identified by precipitation of ABHD17B-FLAG and GFP-FLAG followed by mass spectrometry (MS) (Supplementary Data 8). Enrichment of GO categories (Molecular Functions) for the 15 proteins showing the strongest interaction with ABHD17B (String-db.org) is shown. False discovery rate (FDR) was calculated by Benjamini-Hochberg procedure (String-db.org). d The interactions between the 15 proteins showing the strongest interaction with ABHD17B are displayed in addition to ABHD17B. Dark red lines indicate experimentally-determined interactions (String-db.org). e HSCs were transfected with lentivirus to express GFP-FLAG or ABHD17B-FLAG for 48 h before anti-FLAG precipitation followed by probing for MYO1B (top) and FLAG expression (bottom). Two precipitations were performed for each condition (#1 and #2) and compared to total lysates (L). Data are representative of two independent experiments. f Relative mRNA expression quantified by qRT-PCR in primary human HSCs (Donor 4) treated witih non-targeting siRNAs (NTC si5) and pooled siRNAs targeting MYO1B. Error bars represent mean ± SEM (n = 8 biological replicates). 2-tailed unpaired t-test. Data are representative of three independent experiments. g Wound healing assay was performed in HSCs (donor 4) transfected with siRNAs targeting MYO1B (green) and ABHD17B (blue). Normalized wound width was calculated at the indicated time points from three individual scratches. Error bars represent mean ± SEM (n = 3 biological replicates). One-way ANOVA test. Data are representative of two independent experiments. Source data are provided as Supplementary Data and a Source Data file.

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