Fig. 4

Inhibition of autophagy via PI3K/AKT/mTOR signaling pathway activation causes manchette defects in Actl7a-KO mice. (A-C) The proteome data identified differentially expressed genes (DEGs) in testes from WT and Actl7a-KO male mice. (A) DEGs are represented as a volcano plot. Red dots represent significantly up-regulated genes (n = 33), and blue dots represent significantly down-regulated DEGs (n = 69) compared with WT control. X-axis: log2 fold change (log2 FC), Y-axis: –log10 (P-value). (B) Heatmap analysis of DEGs shows the hierarchical clustering between two groups. Each row represents a single gene. Red represents high gene expression, and blue represents low gene expression. (C) KEGG enrichment analysis displays the top ten pathways of DEGs. X-axis: gene radio, Y-axis: KEGG pathways. The dot size is proportional to gene count, and P-value is indicated by color. (D) WB using PI3K, p-PI3K, AKT, p-AKT, mTOR, and p-mTOR antibodies on protein extracts from WT and Actl7a-KO testes. (E) WB using LC3B-I/II, SQSTM1/p62 and PDLIM1 antibodies on protein extracts from WT and Actl7a-KO testes. β-actin was used as a loading control. IHC using (F) LC3B-I/II, (G) SQSTM1/p62, and (H) PDLIM1 antibodies on WT, and Actl7a-KO testes sections. Scale bars: 50 μm