Journal: Investigative Ophthalmology & Visual Science

Article Title: AIF-1 Drives Corneal Neovascularization by Promoting Inflammatory Macrophage Activation via the MAPK and PI3K/AKT/mTOR Signaling Pathways

doi: 10.1167/iovs.67.2.22

Figure Lengend Snippet: AIF-1 siRNA inhibited the phosphorylation of P38, ERK1/2, JNK, PI3K, AKT, and mTOR proteins in corneal tissues following alkali burn. ( A ) WB analysis of p-P38, p-ERK1/2, and p-JNK expression levels in CNV corneas. ( B – D ) Quantitative analysis of the WB bands ( n = 3). ( E ) The protein expression level of p-PI3K, p-AKT, and p-mTOR in CNV corneas. ( F-H ) Quantitative analysis of the WB bands ( n = 3). β-Actin served as an internal control. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Data are presented as the mean ± SEM.

Article Snippet: The PVDF membrane was incubated overnight at 4°C with primary antibodies, including AIF-1 (1:1000; Abcam, Cambridge, UK), VEGFA (1:1000; CST, Danvers, MA, USA), CD86 (1:1000; Abcam), TNF-α (1:1000; ABclonal, Wuhan, China), IL-1β (1:2000; ABclonal), IL-6 (1:1000; CST), CD31 (1:1000; R&D Systems, Minneapolis, MN, USA), proliferating cell nuclear antigen (PCNA, 1:1000; ABclonal), p-ERK1/2 (1:1500; Proteintech, Wuhan, China), ERK1/2 (1:1500; Proteintech), P38 (1:1000; CST), p-P38 (1:1000; CST), p-JNK (1:1000; CST), JNK (1:1000; CST), p-PI3K (1:500; Affinity Biosciences, Nanjing, China), PI3K (1:1000; Proteintech), p-AKT (1:3000; Proteintech), AKT (1:1000; Proteintech), p-mTOR (1:5000; Proteintech), mTOR (1:1000; Proteintech), and β-actin (1:2000; Elabscience, Wuhan, China).

Techniques: Phospho-proteomics, Expressing, Control

Journal: Molecular Neurobiology

Article Title: Neuropeptide S Protects Dopaminergic Neurons in a Paraquat-Induced Parkinson’s Model Using SH-SY5Y Cells

doi: 10.1007/s12035-025-05401-7

Figure Lengend Snippet: ERK1/2 ( A ) and Nrf2 ( B ) signaling pathways were evaluated in d-SH-SY5Y cells using ELISA assays (* p < 0.05 compared to control, # p < 0.05 compared to NPS, $ p < 0.05 compared to Paraquat, ϕ p < 0.05 compared to NPS + Paraquat-treated group) ( n = 6)

Article Snippet: Subsequently, membranes were incubated overnight at 4 °C with primary antibodies against p-ERK1/2 (1:1000, AF1015, Affinity), ERK1/2 (1:1000, AF0155, Affinity), and Nrf2 (1:500, NB100-91897, Novus Biologicals).

Techniques: Protein-Protein interactions, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecular Neurobiology

Article Title: Neuropeptide S Protects Dopaminergic Neurons in a Paraquat-Induced Parkinson’s Model Using SH-SY5Y Cells

doi: 10.1007/s12035-025-05401-7

Figure Lengend Snippet: A . pERK1/2:ERK1/2 protein ratios, B . Nrf2 protein levels, C . Blotting image of ERK1/2, p-ERK1/2, Nrf2, and β-actin in d-SH-SY5Y cells. (* p < 0.05, compared to control, # p < 0.05 compared to NPS, $ p < 0.05 compared to Paraquat, ϕ p < 0.05 compared to NPS + Paraquat-treated group) ( n = 3)

Article Snippet: Subsequently, membranes were incubated overnight at 4 °C with primary antibodies against p-ERK1/2 (1:1000, AF1015, Affinity), ERK1/2 (1:1000, AF0155, Affinity), and Nrf2 (1:500, NB100-91897, Novus Biologicals).

Techniques: Control

Journal: Cosmetics

Article Title: Melanogenesis Effect of 7-acetoxy-4-methylcoumarin in B16F10 Melanoma Cells

doi: 10.3390/cosmetics7040094

Figure Lengend Snippet: Figure 7. Effect of 7A4MC on the phosphorylation of p-JNK, p-p38, p-ERK. B16F10 cells were treated with 7A4MC at different concentrations for 4 h. (a) Results of Western blot and protein level of p-JNK, (b) p-p38, and (c) p-ERK. Results are expressed as a percentage of the control. The data represent the mean ± SD of triplicate experiments. * p < 0.05, ** p < 0.01, *** p < 0.005. p: Phosphorylated, t: Total.

Article Snippet: Twenty micrograms of total protein samples were resolved by 10% SDS-PAGE gel and transferred onto polyvinylidene difluoride (PVDF) membranes (Bio-rad, Hercules, CA, USA) at 150 V and 2 h. Membranes were blocked with 5% skim milk at 2 h and incubated with primary antibodies, including MITF antibody (1:500), TRP-1 antibody (1:5000), TRP-2 antibody (1:500), TYR antibody (1:500), Phospho-p44/42 MAPK (Erk1/2)(Thr202/Tyr204) antibody (1:1000), Phospho-p38 MAP Kinase (Thr180/Tyr182) antibody (1:500), Phospho-SAPK/JNK (Thr183/Tyr185) antibody (1:500), p44/42 MAPK (Erk1/2) antibody (1:1000), p38 MAPK antibody (1:500), SAPK/JNK antibody (1:500), P-AKT rabbit antibody (1:1000), (all the antibodies were from Cell signaling, USA) at 4 °C overnight, followed by incubation with HRP Anti-Rabbit IgG secondary antibody (1:10,000) (H&L, Rockland Immunochemicals, Inc., USA) at 25 °C for 1 h. Proteins were detected using an ECL kit (Bio-rad, Hercules, CA, USA) and measured using an image reader (LAS-4000, FUJIFILM, Tokyo, Japan).

Techniques: Phospho-proteomics, Western Blot, Control