Journal: Frontiers in genetics

Article Title: Arbutin Protects Retinal Pigment Epithelium Against Oxidative Stress by Modulating SIRT1/FOXO3a/PGC-1α/β Pathway.

doi: 10.3389/fgene.2022.922807

Figure Lengend Snippet: FIGURE 4 | Arbutin exerted protective effects via the SIRT1/FOXO3A/PGC-1α/β and NF-κB/p65 signaling pathway. (A) qRT-PCR was used to measure transcript levels of the SIRT1/FOXO3a/PGC-1α/β pathway and NF-κB/p65 genes. TBHP decreased the expression of SIRT1, FOXO3a, and PGC-1α/β and increased the expression of NF-κB/p65, whereas mRNA levels in the groups that were pretreated with Arbutin showed reversed trend. (B) western blots were conducted to detect the proteins level of SIRT1, FOXO3a, PGC-1α/β, p-ERK, and NFKB1/P65. (C) imageJ was used to analyze the relative expression level of the proteins mentioned above (*p < 0.05, **p < 0.01, ***p < 0.001, n = 3, bars represent SD).

Article Snippet: The primary antibodies were as follows: SIRT1 (CST, New York, United States, #9475, 1:1000), p44/ 42 MAPK (Erk1/2) (CST, New York, United States, #9102, 1: 1000), FOXO3 (Boster Biological Technology, United States, Pleasanton, PB9196, 1:1000), PPARGC1A (Boster Biological Technology, United States, Pleasanton, BM4898, 1:1000), PPARGC1B (Boster Biological Technology, United States, Pleasanton, A02933-1, 1:1000), NF-κB1 p105/p50(CST, New York, United States, #13586 1:1000), and Phospho-NF-κB p65 (Ser536) (CST, New York, United States, #3033 1:1000).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Frontiers in genetics

Article Title: Arbutin Protects Retinal Pigment Epithelium Against Oxidative Stress by Modulating SIRT1/FOXO3a/PGC-1α/β Pathway.

doi: 10.3389/fgene.2022.922807

Figure Lengend Snippet: FIGURE 5 | Sirtinol diminished the capability of Arbutin to assist ARPE-19 cells to defend against oxidative stress. (A) (B) flow cytometric analysis showed that cells treated with only sirtinol, TBHP, cotreated with sirtinol, and TBHP displayed decreased cellular viability. However, sirtinol conduction diminished the protective capacity of Arbutin. (C) ARPE-19 cells were seeded in a 24-well plate and applied wounds at the confluence of 80%. The cells were pretreated with or without Arbutin and then subjected to TBHP (350 µM); meanwhile, cells in certain groups were incubated with sirtinol. Photos were taken at different time points post distinct treatments. (D) fluorescence images observed that ARPE-19 treated with Arbutin while subjected to sirtinol and then exposed to TBHP was unable to recuperate ΔΨm. (E) with sirtinol administration, the protein levels of the SIRT1/FOXO3a/PGC-1α/β pathway decreased in the presence of Arbutin (*p < 0.05, **p < 0.01, ***p < 0.001, n = 3, bars represent SD).

Article Snippet: The primary antibodies were as follows: SIRT1 (CST, New York, United States, #9475, 1:1000), p44/ 42 MAPK (Erk1/2) (CST, New York, United States, #9102, 1: 1000), FOXO3 (Boster Biological Technology, United States, Pleasanton, PB9196, 1:1000), PPARGC1A (Boster Biological Technology, United States, Pleasanton, BM4898, 1:1000), PPARGC1B (Boster Biological Technology, United States, Pleasanton, A02933-1, 1:1000), NF-κB1 p105/p50(CST, New York, United States, #13586 1:1000), and Phospho-NF-κB p65 (Ser536) (CST, New York, United States, #3033 1:1000).

Techniques: Incubation