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. 2015 Jul 1;128(13):2319–2329. doi: 10.1242/jcs.167049

Fig. 2.

Fig. 2.

Differing effects of PDK knockdown on phosphorylation of serine residues on PDHE1α. (A) Control and PDK knockdown (KD) oocytes were stained with anti-pSer293-PDHE1α antibody (green) and counterstained with propidium iodide to label chromosomes (red). (B) Quantification of pSer293-PDHE1α fluorescence shown in A. (C) Control and PDK-KD oocytes were labeled with anti-pSer300-PDHE1α antibody (green) and counterstained with propidium iodide (red). (D) Quantification of pSer300-PDHE1α fluorescence shown in C. (E) Control and PDK-KD oocytes were labeled with anti-pSer232-PDHE1α antibody (green) and counterstained with propidium iodide (red). (F) Quantification of pSer232-PDHE1α fluorescence in E. Arrows in A, C and E point to chromosome congression failure in PDK1-KD and PDK2-KD oocytes. Yellow arrowheads in E show the pSer232-PDH signal in control metaphase oocytes. Representative confocal sections are shown in A, C and E. For B, D and F, results are mean±s.d. At least 60 oocytes for each group were analyzed, and the experiments were conducted three times. *P<0.05 versus controls. (G) Schematic representation of the PDK paralog phosphorylation site specificity for oocyte PDHE1α. Scale bars: 20 µm.