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Figure 4 | Cilia

Figure 4

From: Primary cilia and aberrant cell signaling in epithelial ovarian cancer

Figure 4

wt and cancer OSE cells enter growth arrest upon serum depletion. Analysis of known cell cycle markers in sub-confluent cultures with serum (0 hour) and confluent cultures serum depleted for 48 and/or 72 hours. A, B) WB analysis showing protein expression of phosphorylated retinoblastoma protein (p-RB) in wt and cancer OSE cells and quantification of p-RB levels in serum depleted cultures relative to cultures with serum and with respect to the loading controls (β-actin). C, D) WB analysis showing protein expression of proliferating cell nuclear antigen (PCNA) in wt and cancer OSE cells and quantification of PCNA levels in serum depleted cultures relative to cultures with serum and with respect to the loading controls (α-tub). E-G) IFM analysis of wt and cancer OSE cells using anti-acetylated α-tubulin (Acet.tub) antibody to detect primary cilia (arrows), and anti-Ki67 antibody to visualize Ki67 expression of cycling cells. Nuclei were visualized with DAPI. Note that most cancer cells enter growth arrest upon serum starvation, as judged by the lack of nuclear Ki67 staining. DAPI, 4′,6-diamidino-2-phenylindole; IFM, immunofluorescence microscopy; OSE, ovarian surface epithelium; WB, western blot; wt, wild type.

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