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Fig. 4. P4H1 is expressed in Dictyostelium tip cells. (A-D) Normal (Ax3) cells, transfected with phyA::RFP as a reporter for P4H1 expression, were developed to the tipped aggregate (A,B), slug (C) or pre-culmination stage (D) under ambient conditions, placed under a coverslip and photographed for RFP fluorescence. A and B are different planes from the same aggregate. Although not all cells were fluorescent, fluorescence was detected throughout the structures including the tip (denoted by asterisk). (E,F) Tips (Pst) and central pre-spore (Psp) regions were dissected from normal (strain NC-4) slugs, and 67 µg of S17 fraction protein (lanes d,e) was analyzed by western blotting for P4H1 using anti-P4H1, or for Skp1 using mAb 4E10. Surrounding lanes contain 200 µg (b,g) or 67 µg (c,f) of S100 protein from stationary phase Ax3 (normal) cells (b,c) or P4H1a^- cells (f,g), or M_r standards (a,h). Positions of hydroxylated/glycosylated and unmodified Skp1 are shown. Both panels are from the same blot, and are representative of two experiments.

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