Fig. 7
The effects of NRG1 on the CoCl2-induced protein levels of cleaved caspase-3 and Bcl-XL. a Representative immunofluorescence image of cells after treatment with 100 µM CoCl2 in the presence or absence of 10 nM NRG1 for 24 h. Cells were fixed and immunostained with anti-cleaved caspase-3 (red) and counterstained with DAPI (blue). Cleaved caspase-3 staining was significantly higher in CoCl2-only treated cells than in control cells. NRG1 protected against CoCl2-induced expression of cleaved caspase-3. b The ratio of cleaved caspase-3-positive cells to the total number of cells (n = 6; **P < 0.01 versus the control group; #P < 0.05 versus the CoCl2 alone group). Scale bars, 20 μm. c Protein expression of Bcl-XL, cleaved caspase-3, caspase-3 and β-actin was analyzed by western blotting. Representative immunoblots showing SH-SY5Y cells treated with 100 µM CoCl2 and either PBS or NRG1 (5 nM or 10 nM) for 36 h. d Quantitative analysis of the data in a. Treatment with CoCl2 significantly increased the expression of cleaved caspase-3. NRG1 attenuated the increase in cleaved caspase-3 expression, as shown by the densitometric values (n = 6; **P < 0.01 versus the control group; #P < 0.05, # #P < 0.01 versus the CoCl2 alone group). e Quantitative analysis of the data in a. Treatment with CoCl2 significantly decreased the expression of Bcl-XL in SH-SY5Y cells. NRG1 inhibited the reduction in Bcl-XL expression, as shown by the densitometric values (n = 6; *P < 0.05 versus the control group; #P < 0.05 versus the CoCl2 alone group). f Protein expression of myc-tag, cleaved caspase-3, caspase-3, and β-actin was analysed after EAAC1-myc transfection. g Representative immunoblots of SH-SY5Y cells treated with 100 µM CoCl2 and either PBS or NRG1 (5 nM) for 36 h. Quantitative analysis of the data in f (Mock: n = 6; ***P < 0.001 versus the control group; ##P < 0.01 versus the CoCl2 alone group: EAAC1-myc: n = 6; ***P < 0.001 versus the control group; ###P < 0.001 versus the CoCl2 alone group). EAAC1 transfection reduced CoCl2-induced cleaved caspase-3 (n = 6; +++P < 0.001)