DEV032243 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. qRT-PCR validation of differential gene expression between Thy1⁺ and Thy1-depleted testis cell fractions measured by microarray analysis. (A) Bcl6b, (B) Gfra1 and (C) Lhx1. Data are the mean±s.e.m. for three different replicate samples.

• Supplemental Figure S2 -

  Fig. S2. Flow cytometric analysis of Thy1 expression by F4/80⁺ macrophages in testes of 8-dpp mouse pups. (A) Representative dot plot of the 8-dpp pup testis cell population stained with isotype control antibody (left) and F4/80-specific antibody (right). F4/80⁺ macrophages were identified in region R1. (B) Evaluation of Thy1 expression by F4/80⁺ pup testis cells in region R1. Black line (white fill), Thy1-specific antibody. Red fill, isotype control immunoglobulin.

• Supplemental Figure S3 -

  Fig. S3. Negative control immunofluorescence evaluation. Cross-sections were incubated with normal rabbit IgG (A) or normal sheep IgG (B) as the primary antibody. Secondary detection involved incubation with Alexa488-conjugated antibody against rabbit or sheep IgG. DAPI staining was used to visualize cell nuclei. Scale bars: 50 µm.

• Supplemental Figure S4 -

  Fig. S4. Examination by RT-PCR of Csf1 expression in MACS isolated Thy1⁺ and Thy1-depleted testis cell populations from pup testes. (A) Csf1 expression. (B) Gapdh expression. MW, 100 bp DNA step ladder. Negative control is without cDNA template.

• Supplemental Table S1 -

  Table S1. Differential gene expression in Thy1⁺ compared with Thy1-depleted testis cell populations. Genes expressed 10-fold or greater in the Thy1⁺ fraction were filtered from the total differential gene expression list.