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. 2018 Jul 13;26(5):812–825. doi: 10.1038/s41418-018-0153-0

Fig. 1.

Fig. 1

CTSS modulates BRCA1 protein expression. a CTSS protease activity was detected using MCF7 cells after exposure to 10 Gy (IR) at indicated time points. *p < 0.05 vs. untreated control cells (Student’s t-test). b MCF7 cells were irradiated with 1 Gy and 10 Gy and after indicated time points, Western blotting was performed on the cells. Protein levels were quantified using Image J software, and data are expressed as the fold change relative to the negative control. c Western blot analysis of CRISPR-Cas9-Control (Con) and -CTSS KO MCF7 cells in the presence or absence of 10 Gy radiation. d MCF7 cells transfected with WT-CTSS, si-RNA, or mutant type of CTSS (C25A; active site Cys25 mutated to Ala). e Western blotting of cytosolic and nuclear fractions from MCF7 cells was performed. Fraction purity and equal loading were assessed by Western blots for Lamin B and GAPDH. f MCF7 cells were irradiated (10 Gy) for 2 h and stained with Flag antibody for BRCA1 (green, g), V5 antibody for CTSS (red, R), and DAPI. Cells were analyzed using immunofluorescence microscopy. Quantification of BRCA1 staining was performed by dividing into V5 positive and negative MCF7 cells. The error bars represent S.D. Data are expressed as the fold change relative to the control. *p < 0.05 (Student’s t-test)