Fig. 6: GA DPRs induces TDP-43 cytosolic retention and causes severe damage to the mouse cortical neurons. | Communications Chemistry

Fig. 6: GA DPRs induces TDP-43 cytosolic retention and causes severe damage to the mouse cortical neurons.

From: Nanoscopic investigation of C9orf72 poly-GA oligomers on nuclear membrane disruption by a photoinducible platform

Fig. 6

a Representive immunofluorescence of endogenous TDP-43 (Green) in neuroblastoma SH-SY5Y after ADP-1 treatment (1 μM), photoinitiated, and then incubated for 24 h. Yellow arrows indicate the cells with TDP-43 cytosolic retention. Scale bars indicate 10 µm. b Quantification analysis of SH-SY5Y cells with TDP-43 cytosolic retention. Three biological replicates were carried out (r = 3). More than 32 cells were counted and analyzed in each groups (n ≥ 32). Mean and standard deviation for +ADP-1 +UV = 57.2 ± 6.5; +ADP-1 −UV = 20.4 ± 5.4; −ADP-1 +UV = 18.6 ± 7.3; −ADP-1 −UV = 17.9 ± 6.3. *** indicates statistical significance where p value < 0.001 (analyzed by two-sided Welch’s T test with Bonferroni correction, test statistic in group comparison betwen +ADP-1 +UV and −ADP-1 −UV: p value = 0.0005, degree of freedom = 4, t value = 2.78; group compariosion betwen +ADP-1 −UV and −ADP-1 –UV: p value = 0.21, degree of freedom = 4, t value = 2.77; group compariosion betwen −ADP-1 +UV and −ADP-1 –UV: p value = 0.30, degree of freedom = 4, t value = 2.77), and n.s. indicates not significant. c Representative immunofluorescence images of 21DIV mouse cortical neurons stained with antibody against β-III-tubulin (red) and TDP-43 (green) treated with or without 1 μM ADP-1 and/or 2 min of UV irradiation. The mouse cortical neurons were further incubated for 24 h after ADP-1 treatment and/or UV irradiation. The DNA counter stain DAPI (blue) was included to identify the location of the nucleus. Scale bar indicates 10 µm. d Quantification analysis of degeneration area in 21DIV mouse cortical neurons. The degeneration area percentage (%) of neuron is quantified as fragemnted neurite area/total nerutite area. Three biological replicates were carried out (r = 3). Mean and standard deviation for +ADP-1 +UV = 36.8 ± 8.8; +ADP-1 −UV = 14.5 ± 5.5; −ADP-1 +UV = 14.4 ± 3.9; −ADP-1 −UV = 12.9 ± 5.0. ** indicates statistical significance where p value < 0.01 (analyzed by two-sided Welch’s T test with Bonferroni correction, test statistic in group compariosion betwen +ADP-1 +UV and −ADP-1 −UV: p value = 0.0086, degree of freedom = 3, t value = 3.18; group compariosion betwen +ADP-1 −UV and −ADP-1 –UV: p value = 0.24, degree of freedom = 4, t value = 2.77; group compariosion betwen −ADP-1 +UV and −ADP-1 –UV: p value = 0.234, degree of freedom = 4, t value = 2.77), and n.s. indicates not significant. e Quantification analysis nuclear-to-cytoplasmic ratio of TDP-43 in mice cortical neurons. Three biological replicates were carried out (r = 3) and counted neuron number (n) in each group: +ADP-1 +UV = 86; +ADP-1 −UV = 55; −ADP-1 +UV = 52; −ADP-1 −UV = 56. Mean and standard deviation for +ADP-1 +UV = 1.75 ± 0.05; +ADP-1 −UV = 2.62 ± 0.42; −ADP-1 +UV = 2.49 ± 0.79; −ADP-1 −UV = 2.66 ± 0.19. ** indicates statistical significance where p value  < 0.01 (analyzed by two-sided Welch’s T test with Bonferroni correction, test statistic in group compariosion betwen +ADP-1 +UV and −ADP-1 −UV: p value = 0.005, degree of freedom = 2, t value = 4.30; group compariosion betwen +ADP-1 −UV and −ADP-1 –UV: p value = 0.299, degree of freedom = 3, t value = 3.18; group compariosion betwen −ADP-1 +UV and −ADP-1 –UV: p value = 0.248, degree of freedom = 2, t value = 4.30), and n.s. indicates not significant.

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