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Additional file 1: Figure S1. A: Agarose gel images of RT-PCR products of different DCAF genes from mouse (A). RT-PCR was performed on total RNA from different tissues of adult C57BL/6 mouse using gene specific primers for 56 different DCAF subfamily genes as well as housekeeping genes (β-actin and 18S rRNA). The PCR products were separated on 1.5% agarose gel and images were captured using gel imager. gDNA: Genomic DNA; -ve: No template control. Figure S1. B: Agarose gel images of RT-PCR products of different Dcaf genes from human (B). RT-PCR was performed on total RNA from different tissues of adult human using gene specific primers for 52 different DCAF subfamily genes as well as housekeeping genes (18S rRNA). The PCR products were separated on 1.5% agarose gel and images were captured using gel imager. gDNA: Genomic DNA; -ve control: No template control. Figure S2. Heat map of WD40 family genes (A) and DCAF subfamily genes (B) showing differential expression in different populations of purified spermatogenic cells from mouse testes. Heat map of WD40 and DCAF genes generated from Cruz et al’s RNA sequencing dataset (da Cruz et al., 2016). Samples are denoted in rows and genes are denoted in columns. Red and green denote highly and weakly expressed genes, respectively. LZ - leptotene and zygotene; 2C - spermatogonia, secondary spermatocytes and Sertoli cells; PS - pachytene spermatocytes; RS - round spermatids. Figure S3. Heat map of WD40 family genes (A) and DCAF subfamily genes (B) showing differential expression in different populations of purified spermatogenic cells from human testes. Heat map of WD40 and DCAF genes generated from Guo et al’s RNA sequencing dataset (Guo et al., 2018). Samples are denoted in columns and genes are denoted in rows. Red and green denote highly and weakly expressed genes, respectively. 1 – Spermatogonial stem cells (SSCs); 2 – Differentiating spermatogonia; 3 – Early primary spermatocytes; 4 – Late primary spermatocytes; 5 – Round spermatids; 6 – Elongated spermatids; 7 & 8 – Sperm. Figure S4. Pie charts showing the gene ontology (GO) categorization of the human WD40 family genes (A) and DCAF subfamily genes (B) according to biological processes. WD40 and DCAF genes of human were classified into 10 GO categories for biological processes using Panther. Size of the pie slice corresponds to the number of genes in a given GO category. Figure S5. Pie and bar charts representing the gene ontology (GO) categorization of the mouse(A, C, E) and human (B, D, F) WD40 family genes according to biological processes (A, B), cellular components (C, D) and molecular functions (E, F) using DAVID. Figure S6. Pie and bar charts representing the gene ontology (GO) categorization of the mouse (A, C, E) and human (B, D, F) DCAF family genes according to biological processes (A, B), cellular components (C, D) and molecular functions (E, F) using DAVID. |
