wt1  (Proteintech)

Journal: bioRxiv

Article Title: COPS5 is Essential for Sertoli Cell Function and Male Fertility in Mice

doi: 10.64898/2025.12.19.695357

Figure Lengend Snippet: A. Immunofluorescence result of WT1 in control and Cops5 cKO mice. In 2-month-old control and Cops5 cKO mice, as well as 5-month-old control mice, WT1 signal is in the nuclei of Sertoli cells along the basement membrane. However, in 5-month-old Cops5 cKO mice, some WT1 signals were present in cytoplasm with intense expression. In some Sertoli cells, although WT1 was in the nuclei, but the protein detached from the basement membrane and moved toward the lumen; others became sparse and irregularly arranged, and some lost directional alignment. B. Immunofluorescence result of vimentin in control and Cops5 cKO mice. In 2-month-old control and Cops5 cKO mice, as well as 5-month-old control mice, strong vimentin signal was observed in the perinuclear region, and vimentin-positive extensions, projecting toward the lumen. However, in 5-month-old Cops5 cKO mice, some cells maintained a similar pattern. In many cells, only weak, dot-like vimentin signals were observed. C. Immunofluorescence result of DDX4 in control and Cops5 cKO mice. a. Representative images of DDX4 staining in 2 and 5-month-old control and Cops5 cKO mice. Notice that the DDX4 staining pattern was the same in all the samples. However, in the 5-month-old Cops5 cKO mice, the number of DDX4 positive cells was dramatically reduced. b. Statistic analysis of DDX4 positive cells/seminiferous tubule between the control and Cops5 cKO mice. There was no significant difference between 2-month-old control and Cops5 cKO mice. However, in 5-month-old mice, the Cops5 cKO mice had significantly reduced DDX4 positive cells/seminiferous tubule.

Article Snippet: The antibodies used in this study are as follows: Sertoli cell marker WT1 (Proteintech, 12609-1-AP, Dilution ratio: 1:200); germ cell marker DDX4 (Abcam, Cambridge, UK, ab27591, Dilution ratio:1:400); Tight junction Adapters protein ZO-1 (Thermofisher, Carlsbad, CA, USA 33-9100, Dilution ratio: 1:200); Gap junction Integral membrane protein Connexin 43 (Cell Signaling Technology, Danvers, MA, USA, 3512S, Dilution ratio: 1:200); Goat Anti-Mouse IgG H and L (Abcam, Cambridge, UK, ab150113, Dilution ratio: 1:2000); Goat Anti-Rabbit IgG H and L (Abcam, Cambridge, UK, ab150078, Dilution ratio: 1:2000).

Techniques: Immunofluorescence, Control, Membrane, Expressing, Staining

Journal: bioRxiv

Article Title: COPS5 is Essential for Sertoli Cell Function and Male Fertility in Mice

doi: 10.64898/2025.12.19.695357

Figure Lengend Snippet: A. Analysis of ZO-1 staining pattern. ZO-1, a tight junction protein, and a key marker for Sertoli cell function and BTB integrity was analyzed by Immunofluorescence staining. In 2 and 5-month-old control and 2-month-old Cops5 cKO mice, ZO-1 formed continuous, fence-like lines around the base of the seminiferous tubules, clearly separating the basal and adluminal compartments (A). In contrast, in 5-month-old Cops5 cKO mice, the ZO-1 signal was discontinuous, faint, and punctate, indicating a disrupted BTB structure. B. Analysis of Connexin 43 staining pattern. Connexin 43 (Cx43) is a key component of gap junction. In 2 and 5-month-old control and 2-month-old Cops5 cKO mice, Cx43 appeared as distinct, discrete punctate spots at cell-cell contacts near the basement membrane. This signal was greatly reduced or lost in the 5-month-old Cops5 cKO mice.

Article Snippet: The antibodies used in this study are as follows: Sertoli cell marker WT1 (Proteintech, 12609-1-AP, Dilution ratio: 1:200); germ cell marker DDX4 (Abcam, Cambridge, UK, ab27591, Dilution ratio:1:400); Tight junction Adapters protein ZO-1 (Thermofisher, Carlsbad, CA, USA 33-9100, Dilution ratio: 1:200); Gap junction Integral membrane protein Connexin 43 (Cell Signaling Technology, Danvers, MA, USA, 3512S, Dilution ratio: 1:200); Goat Anti-Mouse IgG H and L (Abcam, Cambridge, UK, ab150113, Dilution ratio: 1:2000); Goat Anti-Rabbit IgG H and L (Abcam, Cambridge, UK, ab150078, Dilution ratio: 1:2000).

Techniques: Staining, Marker, Cell Function Assay, Immunofluorescence, Control, Membrane