Figure 6.

ATM-initiated DSB signaling is involved in modulation of the ENZA response by CK1α

(A) Scatterplot together with Spearman’s correlation was applied to evaluate the correlation strength between the duration of ARSI treatment and ATM mRNA expression based on the “SU2C PNAS2019” datasets. A linear regression line helps visualize the correlation. The analysis concerns only those patients whose ARSI treatment status is recorded as “off treatment” = TRUE (n = 50).

(B) Histogram of the FPKM-normalized gene level of ATM of the “SU2C PNAS-2019” samples were plotted to demonstrate the distribution of ATM levels (n = 55). The dashed line indicates the median expression of ATM.

(C) Kaplan-Meier curve together with the log-rank p value was generated to evaluate the association between the possibility of patients continuing on the line of ARSI treatment and the ATM expression level (dichotomized as high versus low with median chosen as the cutoff) (n = 55).

(D) GSEA showing the enrichment of the ATM pathway in non-responsive (NR) patients to ENZA compared with patients responsive (R) to ENZA (dataset from the study reported by Alumkal et al.⁷).

(E–G) Representative images of ATM (E), γh2AX (F), and cleaved caspase 3 (G) IHC staining in LuCaP 35CR PDXs with the indicated treatments. Scale bars, 100 μm (top) and 20 μm (bottom).

(H) In vitro proliferation of 22Rv1 cells expressing sgCtrl or sgCK1α or sgCK1α and sgATM was determined by AquaBluer assay after treatment for 72 h with the indicated concentrations of ENZA. Inset: IB analysis of whole-cell lysates from 22Rv1 cells expressing sgCtrl or sgCK1α or sgCK1α and sgATM.

(I and J) In vitro proliferation of 22Rv1 cells expressing sgCtrl or two sgRNAs against ATM was detected by AquaBluer assay after combined treatment with the indicated concentrations of ENZA and 50 nM BTX-A51 (I) or 10 μM D4476 (J) for 72 h.