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. 2015 Apr 16;46(6):2299–2308. doi: 10.3892/ijo.2015.2963

Figure 5.

Figure 5

Figure 5

Study of checkpoint/repair response in NO3 NS and AC. (A) Expression by immunofluorescence of checkpoint/repair proteins in untreated NO3 NS (a-g) and in cells treated with 100 μM TMZ for 48 h (h-n). (B) The same as above in untreated (a-g) and treated NO3 AC (h-n). (C) The same as above in NO3 AC treated with 100 nM PTX for 72 h (a-g): positivity of the antigens is evident at metaphasis level. Nuclei counterstained with DAPI. All ×200 magnification. (D) Expression by western blotting of p-ATM, p-Chk2, γ-H2AX and Ku70/Ku80 in 4 NS lines (CV10, 010627, CV21 and NO3), untreated and treated with 100 μM TMZ for 48 h. No band for RAD51 was detectable in any lines, not even after TMZ treatment. (E) Expression by immunocytochemistry of γ-H2AX, as indicator of DNA damage, in untreated NO3 NS (a, ×400) and AC (e, ×400) and after 50 μM TMZ for 48 h [(b) and (f) for NS and AC, respectively, ×400], after 5 μM Dox for 48 h [(c) and (g) for NS and AC, respectively, ×400] and after combined treatment with 50 μM TMZ and 5 μM Dox for 48 h [(d) and (h) for NS and AC, ×400 and ×630, respectively]. (F) Levels of checkpoint/repair proteins at 24 (a) and 72 h (b) in NO3 NS, untreated (control) and treated with 100 μM TMZ, 2 μM Dox or 100 nM PTX.