Journal: Molecular Medicine Reports

Article Title: Effects of Tanshinone IIA on the modulation of miR-33a and the SREBP-2/Pcsk9 signaling pathway in hyperlipidemic rats

doi: 10.3892/mmr.2016.5133

Figure Lengend Snippet: Tanshinone IIA regulation of miR-33a, ABCA1 and ABCG1 expression in the liver. (A) RT-qPCR measurement of miR-33a expression in rat liver tissue. (B) RT-qPCR and western blotting analyses of ABCA1 and ABCG1 expression in liver tissue. Data are presented as the mean ± standard deviation. * P<0.05, ** P<0.01. miR, microRNA; ABCA1, ATP-binding cassette transporter A1; ABCG1, ATP-binding cassette transporter G1; RT-qPCR, reverse trancription-polymerase chain reaction; CON, control; HLP, high lipid diet-fed rats; TAN, Tanshinone IIA treatment of high lipid diet-fed rats; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The membranes were then incubated overnight at 37°C with rabbit anti-rat SREBP-2, LDL-R, PCSK9, ABCA1 (cat no. bs-1627R), ABCG1 (cat no. bs-1231R) and GAPDH (No. bs-2188R) primary antibodies (all 1:300; BIOSS), and then with the HRP-conjugated goat anti-rabbit secondary antibody (1:200) for 1 h at 37°C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Binding Assay, Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Effects of Tanshinone IIA on the modulation of miR-33a and the SREBP-2/Pcsk9 signaling pathway in hyperlipidemic rats

doi: 10.3892/mmr.2016.5133

Figure Lengend Snippet: Tanshinone IIA regulation of SREBP-2, Pcsk9 and LDL-R mRNA and protein expression levels in the liver. (A) Reverse transcription-polymerase chain reaction measurement of SREBP-2, Pcsk9 and LDL-R mRNA expression levels in rat liver tissue. (B) Western blotting analysis of SREBP-2, Pcsk9 and LDL-R protein expression levels in the liver. Data are presented as the mean ± standard deviation. * P<0.05, ** P<0.01. SREBP-2, sterol regulatory element-binding protein 2; Pcsk9, proprotein convertase subtilisin/kexin type 9; LDL-R, low-density lipoprotein receptor; CON, control; HLP, high lipid diet-fed rats; TAN, Tanshinone IIA treatment of high lipid diet-fed rats; GAPDH, .glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The membranes were then incubated overnight at 37°C with rabbit anti-rat SREBP-2, LDL-R, PCSK9, ABCA1 (cat no. bs-1627R), ABCG1 (cat no. bs-1231R) and GAPDH (No. bs-2188R) primary antibodies (all 1:300; BIOSS), and then with the HRP-conjugated goat anti-rabbit secondary antibody (1:200) for 1 h at 37°C.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Standard Deviation, Binding 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

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Ginsenoside Rb1 Facilitates Browning by Repressing Wnt/β-Catenin Signaling in 3T3-L1 Adipocytes

doi: 10.12659/MSM.928619

Figure Lengend Snippet: Effect of ginsenoside Rb1 (G-Rb1) on browning treated with Compound 3f. ( A ) Oil Red O staining detected the lipid droplets. Scale bar=100 μm. ( B ) Optical density values at 490 nm. ( C ) Lipid droplets’ diameter of ORS was measured by ImageJ. ( D ) Western blotting results for Ucp-1 and β-catenin, with Gapdh as the internal reference. ( E, F ) ImageJ quantized the gray value of each protein band. ( G ) Immunofluorescence was performed to detect the expression of Ucp-1. Scale bar=50 μm. Data are expressed as mean±SD (n=3). * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: After blocking, the membrane was incubated in Tris-buffered saline with Tween 20 with anti-Gapdh antibody (1: 1000; #bs-13282R, Bioss), anti-Ucp-1 antibody (1: 500; #bs-1925R, Bioss), anti-GSK-3β antibody (1: 1000; #bs-0023R, Bioss), anti-pGSK-3β antibody (Ser9; 1: 1000, #bs-2066R; Bioss), and anti-β-catenin antibody (Ser9; 1: 1000; #bs-1165R, Bioss), respectively, at 4°C overnight.

Techniques: Staining, Western Blot, Immunofluorescence, Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Ginsenoside Rb1 Facilitates Browning by Repressing Wnt/β-Catenin Signaling in 3T3-L1 Adipocytes

doi: 10.12659/MSM.928619

Figure Lengend Snippet: Effect of ginsenoside Rb1 (G-Rb1) on browning treated with SLK2001. ( A ) Oil Red O staining (ORS) detected the lipid droplets. Scale bar=100 μm. ( B ) Optical density values at 490 nm. ( C ) Lipid droplets’ diameter of ORS was measured by ImageJ. ( D ) Western blotting results of Ucp-1 and β-catenin, with Gapdh as the internal reference. ( E, F ) ImageJ quantized the gray value of each protein band. ( G ) Immunofluorescence was performed to detect the expression of Ucp-1. Scale bar=50 μm. Data are expressed as mean±SD (n=3). * P <0.05, ** P <0.01, **** P <0.0001.

Article Snippet: After blocking, the membrane was incubated in Tris-buffered saline with Tween 20 with anti-Gapdh antibody (1: 1000; #bs-13282R, Bioss), anti-Ucp-1 antibody (1: 500; #bs-1925R, Bioss), anti-GSK-3β antibody (1: 1000; #bs-0023R, Bioss), anti-pGSK-3β antibody (Ser9; 1: 1000, #bs-2066R; Bioss), and anti-β-catenin antibody (Ser9; 1: 1000; #bs-1165R, Bioss), respectively, at 4°C overnight.

Techniques: Staining, Western Blot, Immunofluorescence, Expressing