Fig 2: Effects of phlorofucofuroeckol-A (PFF-A) on pigment accumulation in zebrafish. Synchronized larvae were treated with PFF-A for 72 h. The positive control was arbutin for measuring melanin content. (a) Survival rate after PFF-A treatment of the in vivo zebrafish model. (b) Melanin content in zebrafish after treatment with different concentrations of PFF-A. Images of the zebrafish larva’s body captured using a microscope (Lionheart FX, BioTek, 4×). Data are normalized to the a-MSH group and are presented as the mean ± standard deviation (SD), n = 3. ## p < 0.01 vs. a-MSH-nontreated group; * p < 0.05, ** p < 0.01, and **** p < 0.0001 vs. a-MSH-treated group; ns, non-significant.

Fig 3: hVECs support ZIKV replication and viral production. (A) VK2 cells were infected with ZIKV-PR (MOI: 0.5). Uninfected and heat-inactivated ZIKV cells were used as controls. Viral replication was monitored through the presence of the viral envelope (E) protein using an indirect immunofluorescence assay (IFA) with the 4G2 primary antibody and fluorescein isothiocyanate (FITC)-conjugated secondary antibody. DAPI was used to stain the nucleus. (B) VK2 was infected with ZIKV-PR (MOI: 0.5). The time course of viral replication was analyzed using IFA, as mentioned above. (C) VK2 cells were infected with ZIKV-UG (MOI: 0.5). Venus fluorescent protein expression cells were visualized with a BioTek LionHeart FX Automated Microscope. The percentages of cells positive for ZIKV-PR were calculated by dividing the average number of ZIKV-infected cells (GFP signal) by the average number of cells. The calculated percentages are located at the lower right corners of the merged images. The total numbers of VK2E6E7 cells expressing the Venus fluorescent protein through ZIKV-UG were quantified using the BioTek LionHeart FX Automated Microscope.

Fig 4: Experimental diets increase Alzheimer's disease–related pathology detected using immunohistochemical fluorescence detection. Examples of immunohistochemical staining for amyloid oligomers (ACU193 and NU2) and phosphorylated tau (AT8 p-tau) in hippocampus of rabbits that were fed the control diet (top row), a diet enriched with 2% cholesterol (middle row), or a diet enriched with 10% saturated fat and 30% fructose (bottom row). ACU193 and NU2 (left and center, respectively) were imaged using the Lionheart automated microscope. Images collected from a full-slide scan at 10× were stitched together, then regions of interest were thresholded uniformly using ImageJ. Red on the images denotes specific signal above the determined threshold for the set of images. AT8 (right) images were collected using the Leica SP8 confocal microscope at 63×. Data shows an increase in AT8 (green) positive sites in the dentate gyrus of experimental diet animals over signal detected in the control diet animals. Increased p-tau was also seen in other hippocampal regions as well as frontal and temporal cortex (data not shown). Selected slices were washed with Tris-buffered saline (TBS) three times for 10 minutes each, permeabilized with 0.3% Triton X-100 (Sigma) in TBS (working buffer) for 30 minutes at room temperature, then blocked with 10% normal goat serum (NGS) diluted in working buffer for 60 minutes at room temperature. Slices were incubated with ACU193, a humanized monoclonal antibody against AßOs, at 1 µg/mL, or NU2, a mouse monoclonal against AßOs, 25 at 1.5 µg/mL, or AT8, anti-phospho tau (Ser 202, Thr 205) (Invitrogen), at 1:500 diluted in 10% NGS. Slices were incubated at 4°C for 3 days on an orbital shaker. After washing three times for 30 minutes each with working buffer at room temperature, slices were incubated with the AlexaFluor secondary (Invitrogen), anti-human for ACU193 or anti-mouse for AT8 and NU2, diluted 1:2000 in 1% NGS working buffer at 4°C overnight. Slices were washed three times (15 minutes each) in TBS, then mounted on SuperFrost slides. Images were collected using the Leica TCS SP8 confocal microscope or BioTek Lionheart FX microscope

Fig 5: hCPCs pretreated with histochrome show downregulation of pro-apoptotic signals and upregulation of anti-apoptotic signals under the oxidative stress condition. (A) hCPCs were pretreated with histochrome for 24 h, oxidative stress was induced in hCPCs by 1 mM H2O2. Expression of apoptosis signaling-related proteins was determined by western blotting. (B) Immunofluorescence was performed with the DNA damage marker ?H2A.X to quantify DNA damage of hCPCs. Images were captured using a LionHeart FX automated microscope (Biotek, Winooski, VT, USA). *** p < 0.001 versus +H2O2, n = 5. Error bars indicate S.E.M.