Fig. 6: Af-CUT&Tag identified miR-122 involved in early liver regeneration.
a. Evaluate the enrichment levels of transcription factors Yap1 and Taz at the Mir122 loci, indicating the changes pre and post partial hepatectomy. Positive values indicate an increase, while negative values indicate a decrease. b. Integrative Genomics Viewer snapshot show the distribution of normalized reads of H3K27Ac and ATAC-seq at gene loci of miR-122 in liver cells pre- and post-partial hepatectomy. c. RT-qPCR detects the expression changes of miR-122 at 24 h pre and post partial hepatectomy (the left shows the Yap1-HiBiT group mice, and the right shows the Taz-HiBiT group mice). RT-qPCR data are shown as the mean ± SD from three independent replicates (unpaired t-test. *p < 0.05,**p < 0.01, ***p < 0.001). d. Schematic diagram of Rosa26-CAG-LSL-Cas9-tdTomato mice injected with rAAV2/8-type virus carrying sgRNA targeting the Mir122 locus for liver cell-specific knockout of the Mir122 gene. e. Representative Oil Red O staining images from the control and miR-122#KO at 24 h after PHx. Magnified view at 200× of the area highlighted at 100× by the red squares is depicted in the right column. Scale bar, 100 μm. RT-qPCR detects the expression levels of miR-122 in the control and miR-122#KO liver sample (n = 3). RT-qPCR data are shown as the mean ± SD from three independent replicates (unpaired t-test. ****p < 0.0001). f. Representative liver samples that were stained with H&E in the control and miR-122#KO at 24 hous after PHx. All fields are shown at magnification 200×. Arrows are used to designate areas of inflammation in the miR-122#KO at 24 h post PHx. Scale bar, 100 μm. g, h. Liver RNA-seq data from the GSE111805 dataset58 were utilized, comparing miR-122 knockout and control conditions. Limma (3.62.1) was employed to identify differentially expressed genes (FDR ≤ 0.05, |log2FC|  > 1), followed by GSEA for functional enrichment analysis. The results indicate that specific knockout of miR-122 in the liver significantly upregulates (I) or downregulates (J) certain signaling pathways compared to the control group. i, j. Representative liver samples that were stained for Ki-67 and PCNA in the control (n = 6) and miR-122#KO (n = 6) at 24 h after partial hepatectomy. Magnified view at 400× of the area highlighted at 200× by the red squares is depicted in the right column. Scale bar, 100 μm. Data are presented as mean ± SD., and **p < 0.01, ***p < 0.001 compared to control group, based on unpaired t-test.
