## 2 Results
### Proteomic Analysis of Retinal Tissues to Evaluate Alterations upon Anti-DNM1L Ab Application  
To analyze the protein changes induced by anti-DNM1L Ab injection, mass spectrometric (MS) analysis of the whole retinal tissue was performed. A total of 968 proteins were identified in both experimental groups (IgG and DNML1 OS). Of these, 28 were significantly up-regulated, and 21 were significantly down-regulated (student's \(t\)-test, Figure 7).  
Figure 6: Anti-DNM1L Ab injection reduces proteins involved in mitochondrial apoptosis. (**A**) Western blot detection of XIAP, total protein expression of BAD (BAD\({}^{\rm total}\)), phosphorylation of BAD at serine 112 (BADP\({}^{112}\)), and housekeeping protein beta-actin. The presented representative western blot is one of six biological replicates. Densitometric analysis of XIAP (**B**), BAD\({}^{\rm total}\) (**C**), and BAD\({}^{\rm 112}\) (**D**). All calculated values were normalized to beta-actin. The ratio of BAD\({}^{\rm 112}\)/BAD\({}^{\rm total}\) was calculated (**E**). All values are given as mean \(\pm\) SEM calculated from six biological replicates. Statistical analysis was performed using student’s \(t\)-test. Significance level: * = \(p\)\(<\) 0.05, **= \(p\)\(<\) 0.01, ns = not significant. Numbers of animals used: \(n\) = 6 (IgG), \(n\) = 6 DNM1L.  
Clusters of significantly changed proteins can be identified, which might be related to neuronal activity or mitochondrial dynamics. To identify the association of these proteins with DNM1L, a STRING protein-protein interaction network analysis was performed (Figure 8).  
The network showed significantly altered proteins (\(p<0.05\), student's \(t\)-test) identified in the MS analysis. The target protein of this study, DNM1L, was not identified as signifi  
Figure 8: STRING protein interaction network. Analysis of the signaling pathway altered by the anti-DNM1L Ab injection. Search tool for the retrieval of interacting proteins (STRING) shows the signaling pathways of the most significantly changed proteins using the medium confidence score (0.4). The target protein of the study, DNM1L, was added manually to the network.  
Figure 7: Illustration of the up- and down-regulation of significantly altered protein expression identified by mass spectrometric analysis. After sacrifice of the animals at the end of the study, the proteins were isolated from retinal tissue and tryptically digested. The resulting peptides were purified and analyzed by MS. Identification, quantification and normalization of the peptides was performed using MaxQuant. For statistical analysis, a Student’s \(t\)-test was used with a significance level \(p<0.05\). The graph shows the up—(**A**) and down-regulated (**B**) proteins in the antibody-treated group compared to the control group. Numbers of animals used: \(n=7\) (IgGG), \(n=8\) DNM1L.  
cantly different in the present MS analysis and was manually added to determine possible interactions with the significantly changed marker protein.  
The interaction network demonstrated that DNM1L interacts directly with the proteins alpha-synuclein (SCNA), N-ethylmaleimide sensitive fusion proteins (NSF), heat shock proteins 8A and 9A (HSP8A, HSP9A), ADP/ATP translocae 2 (SLC25A5, ANT2), and microtubule-associated protein 1A (MAP1A). These direct protein interactors are connected to further interaction subnetworks. According to their association or function within the cell, they were categorized as mitochondrial-associated, cytoskeleton-associated, or involved in vesicle trafficking (Table 1).