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. 2016 Apr 29;7(8):900–909. doi: 10.7150/jca.14506

Figure 3.

Figure 3

Knockdown of p53 decreases sensitivity to platinum in HCT116. (A) There was no detectable difference in the p53 protein level between Parental and shcontrol cells. Relative quantities of p53 were normalized to GAPDH and diagrammed (right). (western blot) (B) The protein p53 was knocked down by lentiviral delivery of shRNA (shp53), and the efficiency was assayed by western blot. Relative quantities of p53 were normalized to β-actin and diagrammed (right). (C) Immunohistochemical staining of p53 in HCT116 3D cultures. (D) After treatment of CDDP for 24 hours, p53, p-p53 and cleaved caspase 3 in 3D cultures were assayed by western blot. Relative quantities were normalized to their loading control and diagrammed, respectively (right). (E) After treatment of CDDP for 48 hours, p53 and cleaved caspase 3 in 3D cultures were assayed by western blot. Relative quantities were normalized to Histone H3 and diagrammed (right). (F) After treatment of CDDP for 24 hours, p53 and cleaved caspase 3 in 2D cultures were assayed by western blot. Relative quantities were normalized to β-actin and diagrammed (right). (G) After treatment of CDDP for 24 hours, apoptosis was detected with TUNEL stain. (H) Cell viabilities of HCT116 3D cultures were assayed by WST. *:p<0.01.