Mechanism of anti-human antigen D (HuD/ELAVL4) autoantibody-induced enteric neuronal apoptosis in irritable bowel syndrome and its potential for targeted intervention

Figure 3 HuD protein counteracted the anti-HuD autoantibody to alleviate apoptosis in SH-SY5Y cells. A: Western blot analysis showing the effect of HuD plasmid transfection in counteracting HuD autoantibody-induced expression of apoptotic proteins in SH-SY5Y cells ; B: Terminal-deoxynucleotidyl transferase mediated nick end labeling showing the effect of HuD plasmid transfection antagonizing the HuD autoantibody on nuclei cleavage in SH-SY5Y. Neurons are indicated by PGP 9.5; C: Immunofluorescence showing the effect of HuD plasmid transfection in counteracting HuD autoantibody-induced cleaved caspase 3 expression in SH-SY5Y cells. Neurons are indicated by PGP 9.5; D: Statistical analysis of the mean fluorescence intensity per cell using terminal-deoxynucleotidyl transferase mediated nick end labeling immunostaining; E: Statistical analysis of the mean fluorescence intensity per cell using cleaved caspase 3 immunostaining. Data are expressed as mean ± SD. One-way and two-way analysis of variance with Tukey’s multiple comparison test were performed, and n = 3 for each group. Scale bar: 50 μm. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001, and ^dP < 0.0001. DAPI: 4’,6-diamidino-2-phenylindole; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; TUNEL: Terminal-deoxynucleotidyl transferase mediated nick end labeling.