Figure 1: SPAG6 expression in lymphoid cells and tissues and presence of SPAG6 protein in the centrosome.

(A) SPAG6 expression in adult WT mice compared to testis (positive control) and negative control (water). SPAG6 is expressed in primary (thymus, bone marrow) and secondary (spleen, lymph node) lymphoid tissues and T cells. (B) Quantitative Spag6 expression in different lymphoid tissues assessed by qRT-PCR and normalized relative to the 18s housekeeping gene. (C) Dot plots and histograms showing the absolute numbers and percentages of T cell subsets in the WT and Spag6KO thymic tissues. (D) Dot plots and histograms showing the absolute numbers and percentages of B cells (B220) and T cell subsets in the WT and Spag6KO spleens and lymph nodes [*p < 0.05]. (E) HEK293 cells were transfected with Spag6/pcDNA3 plasmid, and the cells were stained for DAPI (blue), SPAG6 (red) and γ-tubulin (green), a component of centrosome. Co-localization of SPAG6 and γ-tubulin is indicated by white arrows. (F) Non-transfected HEK293 cells were stained for DAPI (blue), SPAG6 (red) and γ-tubulin (green). The cells were also imaged using differential interface contrast (DIC) mode (upper left image). Co-localization of endogenous SPAG6 and γ-tubulin is indicated by white arrows. (G) Non-transfected HEK293 cells labeled with mouse anti γ-tubulin only (rabbit anti Spag6 primary Ab omitted) followed by staining with DAPI (blue), anti rabbit IgG (red) and anti mouse IgG (green). No co-localization of endogenous SPAG6 and γ-tubulin can be seen at the white arrows [negative control]. (H) Purified WT T and B cells were labeled with anti SPAG6 (red) and anti γ-tubulin (green). Co-localization of SPAG6 and γ-tubulin is indicated by white arrows. (I) Purified WT T and B cells were labeled with mouse anti γ-tubulin only (rabbit anti SPAG6 primary Ab omitted) followed by staining with DAPI (blue), anti-rabbit IgG (red) and anti-mouse IgG (green). No co-localization of SPAG6 and γ-tubulin can be seen at the white arrows [negative control].