Fig. 1: Genome-wide CRISPR screen in alpha cells.

A Overview of the CRISPR screen and subsequent next-generation sequencing (NGS) of enriched guide RNAs (gRNAs). B Representative images (left) and dose-dependent survival curve (right) of alphaTC6 cells after treatment with TG (0, 10, 50, and 100 nM) for seven days. Scale bar: 400 µm. Four biological replicates cultured in independent wells were examined. Data are presented as mean values ± SEM. C Principal component analysis (PCA) of NGS data. PCA plot showing the variance of each group of samples. D Heatmap showing log 2 count per million (cpm) of enriched gRNAs in the survived cells after the treatment with TG (0, 50, 100, 150 and 250 nM). E Volcano plot (shown in log2 fold change) of enriched gRNAs in the control cells versus survived cells in 250 nM TG. F Gene Ontology Analysis of the top enriched gRNAs in Fig. 1E. For panels (E, F), differential expression analysis was performed using limma, applying a one-sided moderated t test to compare drug-treated (250 nM TG) and control groups for genes with increased expression. P-values were adjusted for multiple comparisons using the Benjamini–Hochberg method to control the false discovery rate. G Interactive Pathway analysis of the top 50 proteins which gRNAs were enriched in the group of 250 nM TG treatment. H Transcription factor (TF) enrichment analysis of the top genes whose gRNAs were enriched in the group receiving 250 nM TG treatment. Enrichment was assessed by a one-sided Fisher’s exact test, with p-values adjusted for multiple comparisons using the Benjamini–Hochberg FDR method. Bar heights indicate –log₁₀(raw P-values). Source data are provided as a Source Data file.