Mer [p Tyr754, p Tyr749, p Tyr753] Antibody NB300-690-0.05mg from Novus Biologicals, a Bio-Techne Brand

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Oncotarget
May 03 2016

Fig 1: Growth arrested cells by the associating with GAS6 and Mer receptor correlate with CSC populations in PCa cells(A) Expression of the cell-cycle specific protein markers corresponding to G1, S, G2, or M phase in Fucci-PC3 cells as evaluated by Western blot. (B) GAS6 or Mer mRNA expression in G1, S, G2, or M phase in Fucci-PC3 cells as quantified by real-time PCR. Data are representative of mean with s.d. (Student's t-test). (C) Immunofluorescence staining of CD133 (red)/CD144 (green) in G1, S, G2, or M phase in Fucci-PC3 cells. Blue, DAPI nuclear stain. Bar = 10 μm. (D) Quantification of Figure 3C. Data are representative of mean with s.d. (Student's t-test). (E, F) Mer mRNA expression in CD133–/CD44– or CD133+/CD44+ in coculture PCa cells (PC3, DU145) with osteoblasts as quantified by real-time PCR. Data are representative of mean with s.d. (Student's t-test). (G) Left panel: Immunofluorescence staining of Mer (red)/GAS6 (green) in PCa cells in bone marrow of a PCa patient. Blue, DAPI nuclear stain. Bar = 50 μm. Right panel: Mer (red)/GAS6 (green) positive cells (white arrows) in the magnification of the white rectangle from left panel. Bar = 20 μm.
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Oncotarget
May 03 2016

Fig 2: GAS6 overexpression increases CSCs through activation of Mer signaling in PCa cells(A, B) Percent of cells expressing the CSC phenotype in coculture of PCaControl or PCaGAS6OE with osteoblasts as quantified by FACS analysis. Data in Figure 5A, 5B are representative of mean with s.d. (Student's t-test). (C) The sphere formation of PCaControl cells or PCaGAS6OE cells in suspended sphere culture conditions following the treatment with a Mer inhibitor, UNC1062. Bar = 50 μm. (D) Quantification of prostatospheres in suspended sphere culture condition. *Denotes p < 0.05 between vehicle vs. a Mer inhibitor, UNC1062 treatments by Student's t-test. (E, F) MTT assays of PCaControl cells or PCaGAS6OE following the treatment with a Mer inhibitor, UNC1062 at 3 day cultures in adherent culture conditions. N.S. denotes no significance (p > 0.05) between vehicle vs. a Mer inhibitor, UNC1062 treatments by Student's t-test. (G, H) % CSC from DTC population in bone marrow at 24 hours after intracardiac injection of PCaControl cells or PCaGAS6OE in SCID mice (n = 5) as quantified by FACS analysis. Data are representative of mean with s.d. (Student's t-test). (I) Left panel: Immunofluorescence staining of p-Mer (red)/GAS6 (green) in PCa cells in bone marrow of a PCa patient. Blue, DAPI nuclearstain. Bar = 50 μm. Right panel: p-Mer (red)/GAS6 (green) positive cells (white arrows) in the magnification of the white rectangle from left panel. Bar = 20 μm.
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Oncotarget
May 03 2016

Fig 3: GAS6 overexpression activates the phosphorylation of Mer signaling in prostate epithelial cells or cancer cells(A) GAS6 mRNA expression as quantified by by real-time PCR in PCaControl cells or PCaGAS6OE cells. (B) GAS6 protein as quantified by ELISA in PCaControl cells or PCaGAS6OE cells. Data in Figure 4A, B are representative of mean with s.d. (Student's t-test). (C, D). PCaControl cells or PCaGAS6OE cells were starved for 24 hours and then exposed to 10% FBS of culture condition for 1 hour and phosphorylation of Mer was evaluated by (C) Western blot or (D) immunofluorescence staining. Bar = 50 μm. (E, F) GAS6 and Mer mRNA expression in the PrEPGAS6+/+ cells or PrEPGAS6−/− cells by real-time PCR. (G) Mer (red)/GAS6 (green) or p-Mer (red)/GAS6 (green) in the PrEPGAS6+/+ cells or PrEPGAS6−/− cells was evaluated by immunofluorescence staining. Bar = 50 μm.