Additional file 1 of Defects of microtubule cytoskeletal organization in NOA human testes
Creators: name: Wu, Xiaolong
name: Yun, Damin
name: Sang, Mengmeng
name: Liu, Jianpeng
name: Zhou, Liwei
name: Shi, Jie
name: Wang, Lingling
name: Bu, Tiao
name: Li, Linxi
name: Huang, YingYing
name: Lin, Dengfeng
name: Sun, Fei
name: Cheng, C. Yan
Publisher: name: figshare
Year: 2022
Subjects: subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: Cancer

description: Additional file 1: Figure S1. Differential expression of genes encoding proteins that regulate MT organization in normal vs. OA and NOA human testes based on analyses of scRNA-Seq datasets. (A) scRNA-Seq datasets from normal vs. OA and NOA human testes were analyzed used Seurat package in R toolkit using Rstudio to obtain Dot plot to identify differentially expressed genes encoding genes pertinent to MT organization. It is noted that some of these proteins overlap with proteins examined in Figures S2-S5. Dot size represents the percent (%) expressed, and color depth represents the degree of expression level from 2 to -1. The cell types were Sertoli cells (SC), peritubular myoid cells (PMC), Leydig cells (LC), testicular macrophages (tM), endothelial cells (EC), mast cells, spermatogonia (SPG), round spermatids (RS), and elongated spermatids (ES). (B) Heatmap that illustrate some Sertoli cell differentially regulated genes involved in MT organization in OA and NOA vs. normal testes. (C) Violin plots of some selected Sertoli cell differentially regulated genes involved in MT organization in OA and NOA vs. normal testes. Figure S2. Differential expression of genes encoding proteins involved in microtubule bundling in normal vs. OA and NOA human testes based on analyses of scRNA-Seq datasets. (A) Dot plot of all genes in different testicular cell types that are involved in MT bundling in normal vs. OA nd NOA testes. Dot size represents the percent (%) expressed, and color depth represents the degree of expression level from 2 to -1. The cell types examined herein can be found in Legend to Figure S1. These MT bundling proteins are also involved in mitosis and meiosis to support sister chromatid separation, but also unique structural function such as ectoplasmic specialization (ES) in the testis to support spermatogenesis, as well as structural MAPs (e.g., MAP1a) that confer MT stabilization. For example, MAP1b and MAP1s are involved in the formation of microtubule bundles in neurons 1, 2, 3. (B) Heatmap that illustrate some Sertoli cell differentially regulated genes involved in MT bundling in normal vs. OA and NOA testes. (C) Violin plots of selected Sertoli cell differentially regulated genes involved in MT bundling in normal vs. OA and NOA testes. Figure S3. Differential expression of genes encoding proteins involved in MT nucleation in normal vs. OA and NOA human testes based on analyses of scRNA-Seq datasets. (A) Dot plot of all genes in different testicular cell types that are involved in MT nucleation in normal vs. OA and NOA testes. Dot size represents the percent (%) expressed, and color depth represents the degree of expression level from 2 to -1. The cell types examined herein can be found in Legend to Figure S1. (B) Heatmap that illustrate some Sertoli cell differentially regulated genes involved in MT nucleation in normal vs. OA and NOA testes. (C) Violin plots of selected Sertoli cell differentially regulated genes involved in MT nucleation in normal vs. OA and NOA testes. Figure S4. Differential expression of genes encoding proteins involved in MT depolymerization in normal vs. OA and NOA human testes based on analyses of scRNASeq datasets. (A) Dot plot of all genes in different testicular cell types that are involved in MT depolymerization in normal vs. OA and NOA testes. Dot size represents the percent (%) expressed, and color depth represents the degree of expression level from 2 to -1. The cell types examined herein can be found in Legend to Figure S1. (B) Heatmap that illustrate some Sertoli cell differentially regulated genes involved in MT depolymerization in normal vs. OA and NOA testes. (C) Violin plots of selected Sertoli cell differentially regulated genes involved in MT depolymerization in normal vs. OA and NOA testes. Figure S5. Differential expression of genes encoding proteins involved in MT-based cargo transport in normal vs. OA and NOA human testes based on analyses of scRNASeq datasets. (A) Dot plot of all genes in different testicular cell types that are involved in MTbased cargo transport (e.g., spermatids, residual bodies, phagosomes) across the seminiferous epithelium in normal vs. OA and NOA testes. Dot size represents the percent (%) expressed, and color depth represents the degree of expression level from 2 to -1. The cell types examined herein can be found in Legend to Figure S1. (B) Heatmap that illustrate some Sertoli cell differentially regulated genes involved in MT-based cargo transport in normal vs. OA and NOA testes. (C) Violin plots of selected Sertoli cell differentially regulated genes involved in MTbased cargo transport in normal vs. OA and NOA testes. Table S1. Patient information to obtain testis biopsy samples for histopathology and immunofluorescence analysis*. Table S2. Sources of scRNA-Seq datasets Table S3. Antibodies used for different experiments in this report*. Table S4. Primers used for qPCR.