Figure 6

CDK1 as a potential DNM2 phosphorylating kinase in mouse testes.

(a) Immunofluorescent staining of CDK1 in WT and DNM2Δ testes (Scale bar = 20 μm). (b) Dual immunofluorescent staining of fixed adult testis sections probing for CDK1 and DNM2 (Scale bar = 50 μm). (c) Isolated SPG, PS and RS were fixed and probed for CDK1 and DNM2. The cells were counterstained with DAPI and visualized using confocal microscopy (Scale bar = 5 μm). (d) PLA (Duolink) for CDK1 and DNM2 on isolated SPG, PS and RS. Red foci (arrows) show areas of detection of protein interaction. Cells are counterstained with DAPI (Scale bar = 20 μm). (e) Immunoblot for CDK1 immunoprecipitation (IP) using DNM2 as bait (band observed at 34 kDa): lysate (lane 1) contains precleared lysate proteins (positive control), DNM2 IP (lane 2) and control lane (lane 3) represents rabbit IgG IP. (f) Immunoblot for reciprocal DNM2 IP (band observed at ~100 kDa): lysate (lane 1) contains precleared lysate proteins (positive control), CDK1 IP (lane 2) and control lane (lane 3) represents eluted proteins for rabbit IgG IP. Dotted boxes highlight that in both IPs target protein was pulled down (absent from control).