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. 2022 Jun 27;23(13):7117. doi: 10.3390/ijms23137117

Figure 3.

Figure 3

Sera of AQP4+NMO patients increase DNA damage response in astrocytes. (a) The expression of tp53 increased significantly in the human astrocytes cultured with sera of NMO patients (2224.3 ± 139.3, n = 4) compared to the HCs (1923.2 ± 150.9, n = 4), as was determined with NanoString nCounter technology, (b) the expression of tp53 increased significantly in the astrocytes cultured with sera of NMO patients (1.6 ± 0.6, n = 15) vs. 0.64 ± 0.4 (n = 12) of the HCs, as was determined using rt-QPCR, (c) time-course experiments of H2AX expression. Mouse primary astrocytes were cultured with human sera (20% of media) for 48 h, followed by evaluation of nuclear H2AX expression using immunofluorescence staining, (d) nuclear expression of H2AX was determined after culture of 48 h: HCs: 10.6 ± 3.4%; NMO: 30.3 ± 2.2%. Data are means ± SD (n = 9 for each group), (e) representative immunofluorescence analysis of primary astrocytes cultured with sera from seropositive NMO patient or HC for 48 h (scale bar = 60 μm). Each assay was repeated independently at least three times. Significance was determined by unpaired two-tailed student’s t-test (p * ≤ 0.05, **** ≤ 0.0001,). Error bars in all graphs represent standard deviation.