Fig. 3

NRG1 attenuated the CoCl₂-induced overexpression of EAAC1 and Tau in SH-SY5Y cells. a Immunofluorescence analysis with anti-EAAC1 and anti-Tau was performed 24 h after 100 µM CoCl₂ treatment in the presence or absence of 10 nM NRG1 in SH-SY5Y cells. The cells were fixed and immunostained with anti-EAAC1 (green) and anti-Tau (red), while DAPI (blue) was used as a counterstain. The outlined image (upper) is enlarged (bottom). Scale bars, 20 µm. b Bar graph summarizing the data from neurons showing EAAC1 fluorescence (n = 8; **P < 0.01 versus the control group; ^#P < 0.05 versus the CoCl₂ alone group). c The fluorescence intensity of Tau was measured in each group (n = 8; **P < 0.01 versus the control group; ^#P < 0.05 versus the CoCl₂ alone group). d Expression of phospho-Tau (Ser202, Thr205) after 24 h of incubation under 100 µM CoCl₂ treatment in the presence or absence of 10 nM NRG1 in SH-SY5Y cells as assessed by immunofluorescence. The cells were fixed and immunostained with anti-phospho-Tau (Ser202, Thr205) (green) and anti-Tau (red). Scale bars, 20 µm. e Immunofluorescence analysis with anti-phospho-Tau (Ser422) (green) and anti-Tau (red) was performed 24 h after 100 µM CoCl₂ treatment in the presence or absence of 10 nM NRG1 in SH-SY5Y cells. Scale bars, 20 µm. f Bar graph summarizing the data from neurons showing phospho-Tau (Ser202, Thr205) fluorescence (n = 5; **P < 0.01 versus the control group; ^##P < 0.01 versus the CoCl₂ alone group). g The fluorescence intensity of phospho-Tau (ser422) was measured in each group (n = 6; **P < 0.01 versus the control group; ^#P < 0.05 versus the CoCl₂ alone group). h Quantification of data in d and e. The fluorescence intensity of Tau was measured in each group (n = 11; ***P < 0.001 versus the control group; ^###P < 0.001 versus the CoCl₂ alone group)