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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: SKP2 promotes breast cancer tumorigenesis and radiation tolerance through PDCD4 ubiquitination

Fig. 4

SCFSKP2 increases the survival of breast cancer after radiation treatment via PDCD4 suppression. a WT and SKP2−/− MEF cells were treated with radiation (6GY), and after 1 h total lysates were prepared and subjected to IB analysis. b, c MCF-7-Con, MCF-7-SKP2 and MCF-7-SKP2 with PDCD4 stable expression cells were untreated or treated with radiation (6GY), followed by clonogenic survival assay and MTT assay (n = 3). d, e MDA-MB-231-Con, MDA-MB-231-sgSKP2 and MDA-MB-231-sgSKP2 with shPDCD4 stable expression cells were treated with radiation (6GY), followed by clonogenic survival assay and MTT assay (n = 3). f MCF-7-Con, MCF-7-SKP2 and MCF-7-SKP2 with PDCD4 stable expression cells were treated with radiation (6GY), and after 24 h induction of apoptosis was determined Annexin V staining. Annexin V-positive cells were analyzed with FACS, and a percent of Annexin V-positive cells was quantified and shown in the four-quadrant diagram. g MDA-MB-231-Con, MDA-MB-231-sgSKP2 and MDA-MB-231-sgSKP2 with shPDCD4 stable expression cells were untreated or treated with radiation (6GY), and after 24 h induction of apoptosis was determined by Annexin V staining. Annexin V-positive cells were analyzed with FACS, and a percent of Annexin V-positive cells was quantified and shown in the four-quadrant diagram. h MCF-7-Con, MCF-7-SKP2 and MCF-7-SKP2 with PDCD4 stable expression cells were untreated or treated with radiation (6GY) after 1 h, γ-H2AX expression was determined by immunostaining (Scale bars, 5 um). i MDA-MB-231-Con, MDA-MB-231-sgSKP2 and MDA-MB-231-sgSKP2 with shPDCD4 stable expression cells were untreated or treated with radiation (6GY). After 1 h, γ-H2AX expression was determined by immunostaining (Scale bars, 5 um). j MDA-MB-231-Con, MDA-MB-231-sgSKP2 and MDA-MB-231-sgSKP2 with shPDCD4 stable expression cells were untreated or treated with radiation (6GY) after 1 h, and total lysates were prepared and subjected to IB analysis. b-e, h-i Data represent the mean ± SEM of three independent experiments. Student’s t-test used: *< 0.05; **< 0.01 

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