Journal: Molecules

Article Title: Anticancer Effects of Ursi Fel Extract and Its Active Compound, Ursodeoxycholic Acid, in FRO Anaplastic Thyroid Cancer Cells

doi: 10.3390/molecules26175309

Figure Lengend Snippet: Effects of UF extract and UDCA on the expression of angiogenic proteins in human FRO anaplastic thyroid cancer cells. The cells were treated with UF or UDCA at indicated concentrations for 48 h. The expression of angiogenic proteins TGF-β ( a ), VEGF ( b ), N-cadherin (N-Cad) ( c ), and SIRT-1(d) in the cells was determined by western blotting. ( A ) β-Actin was used as an internal control. The expression of each protein was calculated relative to β-actin expression. Data are representative of three independent experiments. ( B ) * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. normal cells. N, untreated cells; UF25, 25 μg/mL UF extract treatment; UF50, 50 μg/mL UF extract treatment; UDCA50, 50 μM/mL UDCA treatment; UDCA100, 100 μM/mL UDCA treatment; D, 2 nM/mL docetaxel treatment.

Article Snippet: Anti-vascular endothelial growth factor (VEGF) Ab (sc-146) was procured from Novus Biologicals (E. Briarwood Avenue, CO, USA), and anti-sirtuin-1 (Sirt-1) Ab (bc-0921R) was obtained from Bioss Antibodies Inc. (Woburn, MA, USA).

Techniques: Expressing, Western Blot

Journal: Asian Journal of Andrology

Article Title: The near-infrared dye IR-61 restores erectile function in a streptozotocin-induced diabetes modelvia mitochondrial protection

doi: 10.4103/aja.aja_69_20

Figure Lengend Snippet: IR-61 increased Nrf2 activation and restored SIRT1 and SIRT3 expression in cavernous tissue. (a) Representative images of SIRT1 costaining with α-SMA. Scale bar = 100 μm. (b) Representative images of SIRT3 co-staining with α-SMA and quantitative analysis of the fluorescence intensity of SIRT3. Scale bar = 100 μm ( n = 4 per group). (c) Representative images of Western blot analysis of SIRT1 and SIRT3. (d) Quantitative analysis of the SIRT1 fluorescence intensity ( n = 4 per group). (e) Relative SIRT1 and SIRT3 protein levels in the cavernous tissue ( n = 4–5 each group). (f) Western blot analysis of Nrf2 and HO-1 in the cavernous tissue. (g) Relative protein levels of Nrf2 and HO-1 in the corpus cavernosum ( n = 4 per group). * P < 0.05, the indicated group compared with the control group; ** P < 0.01, the indicated group compared with the control group; *** P < 0.001, the indicated group compared with the control group; # P < 0.05, the indicated group compared with the DM group; ## P < 0.01, the indicated group compared with the DM group; ### P < 0.001, the indicated group compared with the DM group. Nrf2: nuclear factor (erythroid-derived 2)-like 2; SIRT1: sirtuin1; SIRT3: sirtuin3; HO-1: heme oxygenase; α-SMA: α-smooth muscle actin; DM: diabetes mellitus; DAPI: 4',6-diamidino-2-phenylindole.

Article Snippet: For immunohistochemical and immunofluorescence staining, the primary antibodies were rabbit anti-Desmin (1:200; Abcam, ab32362), goat anti-α-SMA (1:200; Abcam, ab21027), mouse anti-Calponin (1:200; Proteintech, 13938-1-AP, Rosemont, IL, USA), mouse anti-SM22α (1:100; Proteintech, 60213-1-Ig), rabbit anti-OPN (1:200; Abcam, ab8448), rabbit anti-Ras homolog gene family, member A (RhoA; 1:100; Proteintech, 10749-1-AP), rabbit anti-Rho-associated, coiled-coil containing protein kinase 1 (ROCK1; 1:100; Proteintech, 21850-1-AP), mouse anti-vimentin (1:200; Abcam, ab20346), rabbit anti-cleaved-caspase-3 (1:100; Abcam, ab13847), rabbit anti-sirtuin1 (SIRT1; 1:200; Proteintech, 13161-1-AP), rabbit anti-sirtuin3 (SIRT3; 1:200; Proteintech, 10099-1-AP), goat anti-NLRP3 (1:200; Abcam, ab4207), rabbit anti-caspase-1 (1:100; Abcam, ab179515), and mouse anti-apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC; 1:100; Santa Cruz Biotechnology, sc-514414, Santa Cruz, CA, USA).

Techniques: Activation Assay, Expressing, Staining, Fluorescence, Western Blot, Control, Derivative Assay

Journal: Immunity, Inflammation and Disease

Article Title: Progranulin deficiency suppresses allergic asthma and enhances efferocytosis via PPAR‐γ/MFG‐E8 regulation in macrophages

doi: 10.1002/iid3.779

Figure Lengend Snippet: Progranulin deficiency led to increased Sirtuin1 expression through peroxisome proliferators activated receptors‐γ and promoted interleukin‐10 production in efferocytes. (A) Western blot analysis of SIRT1 in PMs. The results of densitometric analysis were showed as means ± SD ( n = 3). * p < .05, unpaired t test. (B) PMs were treated with DMSO or PPAR‐γ inhibitor GW9662 and assayed for SIRT1 protein after 24 h. The results of densitometric analysis were showed as means ± SD ( n = 3). * p < .05, *** p < .001, **** p < .0001, unpaired t test. (C) PMs were cocultured with apoptotic cells for 12 h and Il10 levels were detected by q‐PCR. The relative mRNA expression was showed as means ± SD (n = 3). ** p < .01, *** p < .001, unpaired t test. The study was repeated for three times. GW, GW9662; PM, peritoneal macrophage; PPAR‐γ, peroxisome proliferator‐activated receptor gamma; SD, standard deviation; SIRT1, sirtuin 1.

Article Snippet: Antibodies used in our experiments were as follows: Anti‐cleaved caspase 3 (Cat. No 9664, clone 5A1E) and anti‐sirtuin 1 (SIRT1) (Cat. No. 3931 clone D60E1) were obtained from Cell Signaling Technology; anti‐MFG‐E8 (Cat. No. 377356, clone H‐3) was obtained from Santa Cruz; anti‐PPAR‐γ (Cat. No. ET1702‐57 clone JF101‐4) was obtained from HuaAn Biotechnology; anti‐PGRN (Cat. No. NBP1‐32076, polyclonal) was obtained from Novus Biologicals; anti‐GAPDH (Cat. No. 10494‐1‐AP polyclonal) was obtained from Proteintech and anti‐PE‐F4/80 Cat. No. 123109, clone BM8) was obtained from BioLegend.

Techniques: Expressing, Western Blot, Standard Deviation

Journal: Immunity, Inflammation and Disease

Article Title: Progranulin deficiency suppresses allergic asthma and enhances efferocytosis via PPAR‐γ/MFG‐E8 regulation in macrophages

doi: 10.1002/iid3.779

Figure Lengend Snippet: Progranulin deficiency promoted the expression of milk fat globule‐epidermal growth factor 8 by upregulating peroxisome proliferators activated receptors‐γ in lung of asthmatic mice. (A) Protein levels of PPAR‐γ, MFG‐E8 and SIRT1 in mice lungs were analyzed by Western blot. The results of densitometric analysis were showed as means ± SD ( n = 3). * p < .05, *** p < .001, **** p < .0001, unpaired t test. (B) Protocol of establishing OVA‐induced allergic airway diseases model. (C) The total number of leukocytes in BALF. Results were shown as means ± SD. (WT PBS, n = 5, WT OVA, n = 4, PGRN KO PBS, n = 3, PGRN KO OVA, n = 4, PGRN KO OVA + GW9662, n = 4) ** p < .01, Welch's t ‐test. (D) Protein levels of MFG‐E8 in mice lungs was analyzed by Western blot. ( n = 2 or 3). The study was repeated for three times. Alum, aluminum; OVA, ovalbumin; PGRN, Progranulin; SD, standard deviation; WT, wild type.

Article Snippet: Antibodies used in our experiments were as follows: Anti‐cleaved caspase 3 (Cat. No 9664, clone 5A1E) and anti‐sirtuin 1 (SIRT1) (Cat. No. 3931 clone D60E1) were obtained from Cell Signaling Technology; anti‐MFG‐E8 (Cat. No. 377356, clone H‐3) was obtained from Santa Cruz; anti‐PPAR‐γ (Cat. No. ET1702‐57 clone JF101‐4) was obtained from HuaAn Biotechnology; anti‐PGRN (Cat. No. NBP1‐32076, polyclonal) was obtained from Novus Biologicals; anti‐GAPDH (Cat. No. 10494‐1‐AP polyclonal) was obtained from Proteintech and anti‐PE‐F4/80 Cat. No. 123109, clone BM8) was obtained from BioLegend.

Techniques: Expressing, Western Blot, Standard Deviation