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. Author manuscript; available in PMC: 2012 Jun 1.
Published in final edited form as: Nat Med. 2011 Nov 13;17(12):1641–1645. doi: 10.1038/nm.2464

Figure 2. Phospho-serine 338 CRAF is upregulated in mitosis and localizes to mitotic spindles in human cell lines and tumor biopsies.

Figure 2

(a) Immunoblot analysis of human colon carcinoma HCT-116 cells asynchronized and synchronized at pro-metaphase. pS338 refers to phospho-S338 CRAF, pMEK refers to phospho-MEK and pHH3 refers to phospho-histone H3. Data are representative of three independent experiments. (b) Confocal microscopy images of human pancreatic XPA-1 and glioblastoma U251 cells during mitosis, stained for phospho-S338 CRAF (in green), α-tubulin (in red) and DNA (TOPRO-3 in blue). Scale bar, 10 μm. White arrows indicate localization of phospho-S338 CRAF at the mitotic spindle. (c) Immunoblot analysis of γ-tubulin immunoprecipitates from human colon carcinoma HCT-116 cells asynchronized and synchronized at pro-metaphase. Data are representative of three independent experiments. (d) Immunohistochemical staining of phospho-S338 CRAF in tumor biopsies from breast cancer patients. Scale bar, 10 μm. (e) Confocal microscopy images of XPA-1 cells treated with KG5, sorafenib, ZM336372, L779450 or paclitaxel and stained for CRAF (in green), γ-tubulin (in red) and DNA (TOPRO-3 in blue). Scale bar, 10 μm. White arrows indicate localization of CRAF at the spindle pole. White circle indicates the absence of CRAF at the spindle pole.