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Fig. 2 | Journal of Neuroinflammation

Fig. 2

From: Mesencephalic astrocyte-derived neurotrophic factor (MANF) protects against Aβ toxicity via attenuating Aβ-induced endoplasmic reticulum stress

Fig. 2

1–42 induces ER stress and up-regulated MANF expression in SH-SY5Y cells. a Representative images of MANF (red) and BiP (green) immunofluorescence labeling of SH-SY5Y cells incubated with DMEM containing 10% FBS, serum-free DMEM, and DMEM containing Aβ1–42 (10 μM) for 24 h, or DMEM containing tunicamycin (TM) (2.5 μg/ml) for 12 h, respectively. The nuclei were stained with DAPI (blue). The enlarged images show the expression and cellular distribution of MANF and BiP, respectively. Scale bar = 10 μm. b Representative images of MANF (red) and CHOP (green) immunofluorescence labeling of SH-SY5Y cells incubated with 10% FBS, serum-free DMEM, and DMEM containing Aβ1–42 (10 μM) for 24 h, or DMEM containing TM (2.5 μg/ml) for 12 h, respectively. The enlarged images show the expression and cellular distribution of MANF and CHOP, respectively. Scale bar = 10 μm. c Quantitation of MANF, BiP, and CHOP mRNA levels in SH-SY5Y cells treated with DMEM containing 10% FBS, serum-free DMEM, DMEM containing Aβ1–42 (10 μM) for 24 h, or DMEM containing TM (2.5 μg/ml) for 12 h, respectively. GAPDH was used as control. d The protein levels of MANF, BiP, CHOP, and cleaved caspase-3 in SH-SY5Y cells treated as indicated. α-tubulin was used as a loading control. e Quantitation of protein levels normalized to the α-tubulin by densitometry in d. All the quantitative data were presented as mean ± SD of at least three independent experiments.*P < 0.05, **P < 0.01. C-caspase-3 cleaved caspase-3, TM tunicamycin

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