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. 2023 Mar 17;62:102677. doi: 10.1016/j.redox.2023.102677

Fig. 6.

Fig. 6

IR enhances GPX4 expression by increasing the protein stability of GPX4. (a) An immunoblot showing the expression of GPX4 in MDA-MB-231 cells treatment with control or IR with or without CHX. Data are representative of n = 3 biologically independent experiments. (b) An immunoblot showing the expression of GPX4 and Hsc70 in MDA-MB-231 cells (left) or MCF-7 (right) transfected with negative control (NC) or Hsc70 siRNA. Data are representative of n = 3 biologically independent experiments. (c) Endogenous GPX4 (left) or Hsc70 (right) was immunoprecipitated from MDA-MB-231 cells subjected to the indicated treatments for 16 h, followed by immunoblotting using an antibody to GPX4 and Hsc70. IR, 4 Gy. Data are representative of n = 3 biologically independent experiments. (d) Endogenous GPX4 (left) or Hsc70 (right) was immunoprecipitated from MCF-7 cells subjected to the indicated treatments for 16 h, followed by immunoblotting using an antibody to GPX4 and Hsc70. IR, 6 Gy. Data are representative of n = 3 biologically independent experiments. (e) An immunoblot showing the expression of GPX4 in IR-treated MDA-MB-231 cells (top) or MCF-7 cells (bottom) treatment with MK2206 at the indicated concentration and time. Data are representative of n = 3 biologically independent experiments. (f, g) Endogenous Hsc70 was immunoprecipitated from MDA-MB-231 cells (f) or MCF-7 cells (g) subjected to the indicated treatments for 12 h, followed by immunoblotting using an antibody to GPX4 and Hsc70. (f) IR, 4 Gy; MK2206 2.5 μM. (g) IR, 6 Gy; MK2206 2.5 μM. Data are representative of n = 3 biologically independent experiments. (h, i) An immunoblot showing the expression of GPX4 in MDA-MB-231 cells (h) or MCF-7 cells (i) treatment with DMSO or MK2206 with or without CHX. Data are representative of n = 3 biologically independent experiments. (j) Cell clone measurement in the indicated MDA-MB-231 cells treated with the 2 Gy IR with or without 2.5 μM MK2206. (k) Lipid peroxidation measurement in the indicated MDA-MB-231 cells treated with the 6 Gy IR with or without 2.5 μM MK2206. j, k, Data are the mean ± s.d.; n = 3 biologically independent experiments. Statistical analysis was performed using an unpaired two-tailed Student's t-test.