Journal: Neurochemistry international

Article Title: Proteostasis and autophagy disruption by the aging-related VGVAPG hexapeptide - preliminary insights into a potential novel elastin-induced neurodegeneration pathway in an in vitro human cellular neuron model.

doi: 10.1016/j.neuint.2025.105992

Figure Lengend Snippet: Fig. 2. Interaction between VGVAPG-induced SIRT2 and HRD1 ligase HRD1 mRNA expression (n=3) after the treatment of the differentiated SH-SY5Y cells with 10 nM of VGVAPG, 10 µM of AGK2, or in co-treatment for 24h, (A). HRD1 (B) and SIRT2 (C) protein expression (n=3) after the treatment of the cells with 10 nM of VGVAPG, 10 µM of AGK2, or in the co-treatment for 48h. Immunofluorescence staining of Ac-α-tub (AlexaFluor595, red channel)/nuclei (DAPI, blue channel) after the treatment of the differentiated SH-SY5Y cells with 10 nM of VGVAPG for 48h (D). The main hypothesis tested in this paper is presented as scheme E. Visualization of HRD1 expression using immunofluorescence staining anti-HRD1 ligase (AlexaFluor595; red channel)/nuclei (DAPI; blue channel) after the treatment of the differentiated SH-SY5Y cells with 10 nM of VGVAPG for 48h (F). The inset of LUTs and inverted LUTs are shown next to the original image as well as the HRD1-AlexaFluor594 fluorescence intensity level (n=25). Scale bar = 40 µm, Magnification = 400x (ZEISS Plan-Apochromat 40x/1.3 Oil DIC (UV) VIS-IR objective). Means ± SD (standard deviations) denoted as *** are statistically different at p<0.001, compared to the control.

Article Snippet: For the multi-stained experiment (#IF3), after 266 incubation with the secondary antibodies, the slides were blocked for 30 minutes with 5% 267 goat serum, and primary mouse anti-SIRT2 antibodies (Santa Cruz Biotechnology, cat. sc-268 28298, dilution 1:100) were added.

Techniques: Expressing, Immunofluorescence, Staining, Fluorescence, Control

Journal: Neurochemistry international

Article Title: Proteostasis and autophagy disruption by the aging-related VGVAPG hexapeptide - preliminary insights into a potential novel elastin-induced neurodegeneration pathway in an in vitro human cellular neuron model.

doi: 10.1016/j.neuint.2025.105992

Figure Lengend Snippet: Fig. 5. Inhibitory effect of VGVAPG on UPS leads to SIRT2 accumulation and ER stress. Caspase-like (β1 subunit) – A; trypsin-like (β2 subunit) – B; and chymotrypsin-like (β5 subunit) – C proteasome activities (n = 6), along with the expression of ubiquitinated proteins (n = 3) – D, were measured after treating neurons with 10 nM VGVAPG, 10 µM AGK2 alone, or in the co-treatment for 48 h. A schematic summarizing the VGVAPG-induced interactions between specific intracellular components is shown on the right (E). Representative blots are shown on the left. Means ± SD (standard deviations) denoted as *** are statistically different at p<0.001, compared to the control (ANOVA). The GAPDH protein expression was always used as a loading control.

Article Snippet: For the multi-stained experiment (#IF3), after 266 incubation with the secondary antibodies, the slides were blocked for 30 minutes with 5% 267 goat serum, and primary mouse anti-SIRT2 antibodies (Santa Cruz Biotechnology, cat. sc-268 28298, dilution 1:100) were added.

Techniques: Expressing, Control

Journal: Neurochemistry international

Article Title: Proteostasis and autophagy disruption by the aging-related VGVAPG hexapeptide - preliminary insights into a potential novel elastin-induced neurodegeneration pathway in an in vitro human cellular neuron model.

doi: 10.1016/j.neuint.2025.105992

Figure Lengend Snippet: Fig. 6. VGVAPG-induced autophagy correlates with the SIRT2 overexpression as a result of inhibition of UPS. Immunofluorescence staining (A) of SIRT2 (AlexaFluor594; red channel), ATG18 (FITC; green channel), and nuclei (DAPI; blue channel) in the differentiated SH-SY5Y cells treated with 10 nM of VGVAPG, 1 µM of LS- 102, or after the co-treatment for 24h. Quantification of SIRT2 (n=40) (B) and ATG18 (n=40) (C) fluorescence. The representative insets (1 – 4) are shown below, together with the colocalization analysis and its quantification based on Pearson’s correlation (n=3) (D). The white arrows indicate the obvious colocalization. Scale bar = 50 µm. Magnification = 630× (ZEISS Plan-Apochromat 63×/1.40 Oil DIC M27). Means ± SD (standard deviations) denoted as *** are statistically different at p<0.001, compared to the control, while means denoted as # are statistically different between certain groups at p<0.05 (t-test).

Article Snippet: For the multi-stained experiment (#IF3), after 266 incubation with the secondary antibodies, the slides were blocked for 30 minutes with 5% 267 goat serum, and primary mouse anti-SIRT2 antibodies (Santa Cruz Biotechnology, cat. sc-268 28298, dilution 1:100) were added.

Techniques: Over Expression, Inhibition, Immunofluorescence, Staining, Fluorescence, Control