Fig 2: Retinal nerve fiber layer (RNFL) thickness analysis. Optical coherence tomography (OCT) was used to scan the retinal cross-sections in vivo. OCT allowed the acquisition of different retinal layers. For the analysis of the acquired circular B-scans with 12° diameter, a semi-automatic algorithm was used to define the RNFL and calculate its thickness (RNFLT). The Heidelberg Eye Explorer software divided the scan into the main sectors superior (S), temporal (T), inferior (I), and nasal (N). Furthermore, the sectors temporal superior (TS), temporal inferior (TI), nasal inferior (NI), and nasal superior (NS) were included (A). In addition to the regional classification, the schematic illustration also showed the sides of the interventions, episcleral vein occlusion (EVO) and intravitreal injection (IVI), which were performed on the eye during the study (Scheme modified from [22]. The mean RNFLT of the whole B-scan was used for follow-up quantification (B). Further quantification of the RNFLT at week 10 was performed for the individual sectors (C–H). Legend: black (IgG OS), red (DNM1L), light grey (IgG OD), dark grey (IgG OD), ** p < 0.01, *** p < 0.001, ns = not significant; One-way ANOVA, Tukey’s HSD test (post hoc). Numbers of animals used: n = 7 (IgG), n = 8 DNM1L.

Fig 3: 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.

Fig 4: 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 (IgG), n = 8 DNM1L.

Fig 5: Anti-DNM1L Ab injection reduces proteins involved in mitochondrial apoptosis. (A) Western blot detection of XIAP, total protein expression of BAD (BADtotal), phosphorylation of BAD at serine 112 (BADp112), and housekeeping protein beta-actin. The presented representative western blot is one of six biological replicates. Densitometric analysis of XIAP (B), BADtotal (C), and BADp112 (D). All calculated values were normalized to beta-actin. The ratio of BADp112/BADtotal was calculated (E). All values are given as mean ± 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.