Journal: iScience

Article Title: O 6 -methylguanine DNA methyltransferase regulates β-glucan-induced trained immunity of macrophages via farnesoid X receptor and AMPK

doi: 10.1016/j.isci.2023.108733

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Article Snippet: The antibodies were diluted in the buffer according to the the manufacturer’s protocol at the following concentration: rat anti-mMGMT, 1:2000 (R&D Systems, USA); rabbit anti-phospho AKT (Thr308), 1:1000; rabbit anti-AKT, 1:1000; rabbit anti-phospho NF-κB p65, 1:2000; rabbit anti-NF-κB p65, 1:4000; rabbit anti-phospho ERK1/2, 1:2000; rabbit anti-ERK1/2, 1:4000; rabbit anti-phospho p38, 1:2000; rabbit anti-p38, 1:4000; rabbit anti-phospho SAPK-JNK, 1:2000; rabbit anti-SAPK-JNK, 1:4000; rabbit anti-phospho mTOR, 1:1000; rabbit anti-mTOR, 1:1000; rabbit anti-HIF1α, rabbit anti-phospho AMPK alpha, 1:2000; rabbit anti-AMPK alpha, 1:2000; anti-actin antibody, 1:10000; and goat anti-rabbit IgG HRP, 1:4000 (all antibodies were purchased from Cell Signaling Technology, USA); goat anti-rat IgG HRP, 1:4000 (Biolegend, USA); sheep anti-mouse IgG HRP, 1:4000 (Cytiva, USA); rabbit polyclonal anti-GAPDH, 1:4000 (Bio-Rad, USA).The signal was detected by the ECL chemiluminescent detection method.

Techniques: Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Western Blot, Control, Software, SYBR Green Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Inhibition of matrix metalloproteinase expression by selective clearing of senescent dermal fibroblasts attenuates ultraviolet-induced photoaging.

doi: 10.1016/j.biopha.2022.113034

Figure Lengend Snippet: Fig. 2. Selective elimination of senescent cells in UV-irradiated mice skin after treatment with ABT-263 or ABT-737. Hairless mice were exposed to chronic UV irradiation for eight weeks with 100 mJ/cm2 for the first 2 weeks, then 50 mJ/cm2 weekly, then 300 mJ/cm2. Each drug (ABT-263, 5 μM; ABT-737, 10 μM) or vehicle (DMSO) was diluted in PBS (30 μl) and injected intradermally into the dorsal skin for a total of ten doses. (A) Representative images of SA-β-gal staining in mice skin. SA-β-gal (blue color) was detected in 6 µm frozen skin sections from unirradiated control mice injected with vehicle (n = 7, white bars), UVB-irradiated mice injected with vehicle (n = 4, black bars), UVB-irradiated mice injected with ABT-263 (n = 6, blue bars) or ABT-737 (n = 7, red bars). Arrowheads indicate stained cells. (B) Quantitative analysis of the percentage of SA-β-gal positive cells in the indicated mice skin (n = 3–4 mice per group) (C) Western blots and quantification of p21 and p16 protein levels normalized to GAPDH using protein extracts from the skin. (D) p16 and p21 mRNA levels in indicated mice skin, standardized to GAPDH. (E) Representative images of immunofluorescence staining for p16 (red) and DAPI (blue) in the skin of the indicated mice. Right: Quan tification of the percentage of p16-positive cells among DAPI-positive cells in over n = 4 mice per group. Arrowheads indicate stained cells. Data represent means ± SEM, *p < 0.05, * *p < 0.005, * **p < 0.001. p-values were calculated by Student’s t-test. A, Bar = 100 µm; E, Bar = 50 µm.

Article Snippet: The antibodies used included p16 (MA5–17142, Thermo Fisher Scientific; ab189034, Abcam), p21 (556431, BD Pharmingen; ab109199, Abcam), GAPDH (CSB-PA00029A0Rb, Cusabio, Houston, TX, USA), β-actin (CSBPA000350, Cusabio), MMP-1 (Lab Frontier, Seoul, Korea), and type I procollagen (SP1.

Techniques: Irradiation, Injection, Staining, Control, Western Blot, Immunofluorescence