Journal: Aging and Disease

Article Title: Role of the cGAS-STING Pathway in Aging-related Endothelial Dysfunction

doi: 10.14336/AD.2022.0316

Figure Lengend Snippet: Changes in eNOS, p53, p21, p16, cGAS, STING and p-IRF3/IRF3 expression levels in D-GAL-treated HAECs. (A) The eNOS, p53, p21, p16, cGAS, STING, IRF3, p-IRF3 and β-actin protein expression levels were measured in HAECs treated with 0, 5, 10, 15, 20 and 25 g/L D-GAL for 24 h by western blot analysis. The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (C). Data analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. AU indicates arbitrary units. Relative expression is the fold changes relative to the 0 g/L group. n=6, * P <0.05 compared with the 0 g/L group. A concentration of 20 g/L was chosen for use in subsequent experiments. (B) The eNOS, p53, p21, p16, cGAS, STING, IRF3, p-IRF3 and β-actin protein expression levels were measured in HAECs at 0, 6, 12, 24 and 48 h after treatment with 20 g/L D-GAL by western blot analysis. The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (D). Data were analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. AU indicates arbitrary units. Relative expression is the fold changes relative to the 0 h group. n=6, * P <0.05 compared with the 0 h group. A time point of 48 h was chosen for use in subsequent experiments

Article Snippet: RU.521 (Cat# HY-114180), H-151 (Cat# HY-112693) and D-GAL (Cat# HY-N0210) were obtained from MCE (NJ, USA).

Techniques: Expressing, Western Blot, Concentration Assay

Journal: Aging and Disease

Article Title: Role of the cGAS-STING Pathway in Aging-related Endothelial Dysfunction

doi: 10.14336/AD.2022.0316

Figure Lengend Snippet: Effects of RU.521 on eNOS, cell senescence, inflammatory cytokines and NO production in D-GAL-treated HAECs. 3μM RU.521 was used to treat the HAECs. eNOS, p53, p21, p16, cGAS, p-STING, STING, IRF3, p-IRF3 and β-actin expression levels in HAECs from the Control, D-GAL and D-GAL+RU.521 groups were examined by western blot analysis (A). The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (B). mRNA expression of IFNβ (C), Ifit1 (D), Ifit2 (E), Ifit3 (F), MCP-1 (G), IL-1β (H), IL-6 (I) and TNF-α (J) in HAECs from the Control, D-GAL and D-GAL+RU.521 groups was determined by PCR. NO release into the culture medium of HAECs from the Control, D-GAL and D-GAL+RU.521 groups was measured (K). Representative photomicrographs and quantitative analysis of SA-β-gal-positive staining in the Control, D-GAL and D-GAL+RU.521 groups (L and M). Data were analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. AU indicates arbitrary units. Relative expression is the fold changes relative to the Control group. n=6, * P <0.05 compared with the Control group, # P <0.05 compared with the D-GAL group.

Article Snippet: RU.521 (Cat# HY-114180), H-151 (Cat# HY-112693) and D-GAL (Cat# HY-N0210) were obtained from MCE (NJ, USA).

Techniques: Expressing, Control, Western Blot, Staining

Journal: Aging and Disease

Article Title: Role of the cGAS-STING Pathway in Aging-related Endothelial Dysfunction

doi: 10.14336/AD.2022.0316

Figure Lengend Snippet: Effects of siRNA-cGAS on eNOS, cell senescence, inflammatory cytokines and NO production in D-GAL-treated HAECs. cGAS expression in the Control, Negative Control (NC) and si-cGAS groups was measured by western blot analysis 48 h after transfection (A). The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (B). Data were analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. n=6, & P <0.05. eNOS, p53, p21, p16, cGAS, p-STING, STING, IRF3, p-IRF3 and β-actin expression levels in HAECs from the Control, D-GAL and D-GAL+si-cGAS groups were examined by western blot analysis (C). The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (D). mRNA expression of IFNβ (E), Ifit1 (F), Ifit2 (G), Ifit3 (H), MCP-1 (I), IL-1β (J), IL-6 (K) and TNF-α (L) in HAECs from the Control, D-GAL and D-GAL+si-cGAS groups was determined by PCR. NO release into the culture medium of HAECs from the Control, D-GAL and D-GAL+si-cGAS groups was measured (M). Representative photomicrographs and quantitative analysis of SA-β-gal-positive staining in the Control, D-GAL and D-GAL+si-cGAS groups (N and O). Data were analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. AU indicates arbitrary units. Relative expression is the fold changes relative to the Control group. n=6, * P <0.05 compared with the Control group, # P <0.05 compared with the D-GAL group.

Article Snippet: RU.521 (Cat# HY-114180), H-151 (Cat# HY-112693) and D-GAL (Cat# HY-N0210) were obtained from MCE (NJ, USA).

Techniques: Expressing, Control, Negative Control, Western Blot, Transfection, Staining

Journal: Aging and Disease

Article Title: Role of the cGAS-STING Pathway in Aging-related Endothelial Dysfunction

doi: 10.14336/AD.2022.0316

Figure Lengend Snippet: Effects of H-151 on eNOS, cell senescence, inflammatory cytokines and NO production in D-GAL-treated HAECs. H-151 (3 μM) was used to treat HAECs. eNOS, p53, p21, p16, cGAS, STING, IRF3, p-IRF3 and β-actin expression levels in HAECs from the Control, D-GAL and D-GAL+H-151 groups were examined by western blot analysis (A). The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (B). mRNA expression of IFNβ (C), Ifit1 (D), Ifit2 (E), Ifit3 (F), MCP-1 (G), IL-1β (H), IL-6 (I) and TNF-α (J) in HAECs from the Control, D-GAL and D-GAL+H-151 groups was determined by PCR. NO release into the culture medium of HAECs from the Control, D-GAL and D-GAL+H-151 groups was measured (K). Representative photomicrographs and quantitative analysis of SA-β-gal-positive staining in the Control, D-GAL and D-GAL+H-151 groups (L and M). Data were analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. AU indicates arbitrary units. Relative expression is the fold changes relative to the Control group. n=6, * P <0.05 compared with the Control group, # P <0.05 compared with the D-GAL group.

Article Snippet: RU.521 (Cat# HY-114180), H-151 (Cat# HY-112693) and D-GAL (Cat# HY-N0210) were obtained from MCE (NJ, USA).

Techniques: Expressing, Control, Western Blot, Staining

Journal: Aging and Disease

Article Title: Role of the cGAS-STING Pathway in Aging-related Endothelial Dysfunction

doi: 10.14336/AD.2022.0316

Figure Lengend Snippet: Effects of si-STING on eNOS, cell senescence, inflammatory cytokines and NO production in D-GAL-treated HAECs. STING expression in the Control, Negative Control (NC) and si-STING groups was measured 48 h after transfection (A). The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (B). Data analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. n=6, & P <0.05. eNOS, p53, p21, p16, STING, IRF3, p-IRF3 and β-actin protein expression levels in HAECs from the Control, D-GAL and D-GAL+si-STING groups were examined by western blot analysis (C). The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (D). mRNA expression of IFNβ (E), Ifit1 (F), Ifit2 (G), Ifit3 (H), MCP-1 (I), IL-1β (J), IL-6 (K) and TNF-α (L) in HAECs from the Control, D-GAL and D-GAL+si-STING groups was assessed by PCR. NO release in the medium of the Control, D-GAL and D-GAL+si-STING groups was measured (M). Representative photomicrographs and quantitative analysis of SA-β-gal-positive staining in the Control, D-GAL and D-GAL+si-STING groups (N and O). Data analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. AU indicates arbitrary units. Relative expression is the fold changes relative to the Control group. n=6, * P <0.05 compared with the Control group, # P <0.05 compared with the D-GAL group.

Article Snippet: RU.521 (Cat# HY-114180), H-151 (Cat# HY-112693) and D-GAL (Cat# HY-N0210) were obtained from MCE (NJ, USA).

Techniques: Expressing, Control, Negative Control, Transfection, Western Blot, Staining

Journal: Biomolecules

Article Title: The Ensemble of Conformations of Antifreeze Glycoproteins (AFGP8): A Study Using Nuclear Magnetic Resonance Spectroscopy

doi: 10.3390/biom9060235

Figure Lengend Snippet: Sequence-specific chemical shift assignments of antifreeze glycoprotein fraction 8 (AFGP8). The 2D TOCSY NMR spectra of AFGP8-BS (AFGP8 from Boreogadus saida ) ( a ) and AFGP8-TB (AFGP8 from Trematomus borchgrevinki ) ( b ), showing the amide to alpha and methyl proton regions. The TOCSY patterns associated with all the amino acid residues of the primary sequence, except for the alanine residue at position 1 and proline residues at positions 4 and 10 in AFGP8-BS and positions 7 and 13 in AFGP8-TB.

Article Snippet: The disaccharide beta- d -galactosyl-(1,3)-alpha-N-acetyl- d -galactosamine (5 mg, 98% purity, CAS 3554-90-3, Cat No. A152000) was purchased from Toronto Research Chemicals (North York, ON, Canada) and was used as purchased.

Techniques: Sequencing, Residue

Journal: Process Biochemistry

Article Title: A novel cold-active β-d-galactosidase with transglycosylation activity from the Antarctic Arthrobacter sp. 32cB – Gene cloning, purification and characterization

doi: 10.1016/j.procbio.2014.09.018

Figure Lengend Snippet: Fig. 7. TLC of products of alkyl glycosides synthesis catalyzed by Arthrobacter sp. 32cB -d-galactosidase. Reaction mixtures containing lactose and 1-butanol (A), 2-propanol (B), 1-hexanol (C) or cyclohexanol (D) as substrates, and 2 U mL−1 Arthrobacter sp. 32cB -d-galactosidase were incubated at 30 ◦C for 8 h. Lane 1 – d-galactose, lane 2 – d-glucose,

Article Snippet: Materials Isopropyl- -d-thiogalactopyranoside (IPTG) and 5-bromo4-chloro-3-indolyl- -d-galactopyranoside (X-gal) were purchased from Biosynth AG (Switzerland). o-Nitrophenyl-d-galactopyranoside (ONPG), p-nitrophenyl- -d-galactopyranoside (PNPG), p-nitrophenyl- -d-glucopyranoside, p-nitrophenyl- -d-fucopyranoside, p-nitrophenyl- -d-xylopyranoside, p-nitrophenyl- -d-glucouronide, p-nitrophenyl- -d-manno pyranoside, p-nitrophenyl- -d-cellobioside, p-nitrophenyl- - l-arabinopyranoside, p-nitrophenyl- -d-galactopyranoside, p-nitrophenyl- -d-glucopyranoside, l-arabinose, d-xylose, lactulose, dithiothreitol (DTT), ethylenediaminetetraacetic acid sodium salt (EDTA), N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid) (HEPES) and tris(2-carboxyethyl)phosphine (TCEP) were supplied by Sigma (USA).

Techniques: Incubation