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. 2013 Feb 26;4:1508. doi: 10.1038/ncomms2489

Figure 4. Knockdown of PIB5PA results in anchorage-independent growth of melanocytes.

Figure 4

(a) The PIB5PA mRNA expression levels in HEMa-LP melanocytes transduced with the control or PIB5PA shRNA by quantitative PCR (n=3, mean±s.e.m.). (b) Whole-cell lysates from HEMa-LP melanocytes transduced with the control or PIB5PA shRNA were subjected to western blot analysis. (c) Anchorage-independent growth assay of HEMa-LP melanocytes transduced with the control, PIB5PA or PTEN shRNA. Scale bar, 0.5 mm. (d) Quantification of anchorage-independent growth assay of HEMa-LP melanocytes transduced with the control, PIB5PA or PTEN shRNA as shown in c (n=3, mean±s.e.m.; Student’s t-test; *P<0.05). (e) HEMa-LP melanocytes transduced with the control, PIB5PA or PTEN shRNA were counted on days 3, 6 and 9, using an automated cell counter (n=3, mean±s.e.m.). (f) HEMa-LP melanocytes transduced with the PIB5PA shRNA were cointroduced with the control or Akt shRNA. Twenty-four hours later, whole-cell lysates were subjected to western blot analysis. (g) HEMa-LP melanocytes transduced with the PIB5PA shRNA were cointroduced with the control or Akt shRNA. Ninety-six hours later, viable cells were counted in an automated cell counter (n=3, mean±s.e.m.).