Journal: CNS neuroscience & therapeutics

Article Title: Alleviation of Microglia Mediating Hippocampal Neuron Impairments and Depression-Related Behaviors by Urolithin B via the SIRT1-FOXO1 Pathway.

doi: 10.1111/cns.70379

Figure Lengend Snippet: FIGURE 5 | The effects of UB on the ROS and the phosphorylation levels of P65, ERK, P38, JNK, and AMPK of BV2 cells. (A) Experimental flow diagram. After pretreating with UB (0, 10, 20, 40 μmol/L) for 2 h, BV2 cells were treated with LPS (0.5 μg/mL) for 24 h. ROS and RNA extraction were performed. (B and C) Intracellular ROS levels were measured using the DCFH-DA method. Scale bar, 50 μm. (D and E) Western blot was performed to detect the phosphorylation levels of P65, ERK, P38, JNK, and AMPK, the acetylation level of P65, and the expression levels of IDO of BV2 cells. n = 3 per group. #p < 0.05, ##p < 0.01, ###p < 0.001 versus the control group without LPS and UB treatment or the group treated with LPS alone.

Article Snippet: The primary antibodies employed in this study were as follows: rabbit anti- SIRT1 (Sigma- Aldrich, 07- 131), rabbit anti- FOXO1 (CST, 2880S), mouse anti- β- actin (CST, 3700S), rabbit anti- P65 (CST, 8242T), rabbit anti- phospho- P65 (CST, 3031S), ARG1 (BOSTER, A01106), CD206 (BOSTER, A02285- 2), CD86 (BOSTER, A00220- 4), iNOS (BOSTER, BA0362), ERK (CST, 4695), p- ERK (CST, 4370), rabbit anti- P38 (CST, 9212S), phospho- P38 (CST, 9215), JNK (CST, 9251), phospho- JNK (CST, 9252), rabbit anti- AMPK (CST, 5831S), rabbit anti- phospho- AMPK (CST, 2535S), rabbit anti- Bax (CST, 5023T), mouse anti- Bcl- 2 (CST, 15071T), mouse anti- IDO (Proteintech, 66528- 1- Ig), rabbit anti- Ac- FOXO1 (Invitrogen, PA5- 104560), rabbit anti- Ac- P65 (Abcam, AB19870).

Techniques: Phospho-proteomics, RNA Extraction, Western Blot, Expressing, Control

Journal: Journal of the American Heart Association

Article Title: SWAP70 Overexpression Protects Against Pathological Cardiac Hypertrophy in a TAK1‐Dependent Manner

doi: 10.1161/jaha.122.028628

Figure Lengend Snippet: Figure 1. SWAP70 protein level is increased in response to hypertrophic stimuli. A, Immunoblotting and quantitation of SWAP70, ANP, BNP, MYH7/β-MHC protein levels in human heart tissues from normal donors (n=5), HCM heart failure patients (n=5), and DCM heart failure patients (n=7). B, Relative mRNA levels of SWAP70 in human heart tissues from normal donors (n=5), HCM heart failure patients (n=5), and DCM heart failure patients (n=9) (NS: left, P=0.90; right, P=1.00). C, Left, expression of SWAP70 in human tissues from normal donors or heart failure patients arise from HCM or DCM. Right, quantitation results of immunohistochemical staining (n=4). D, Protein levels and quantitation results of SWAP70, ANP, BNP, MYH7 in mice hearts subjected to TAC surgery at the indicated time points (n=6). E, Relative mRNA levels of Swap70 in mice heart tissues after TAC operation at indicated time points (n=5) (NS: left, P=1.00; middle, P=0.11; right, P=0.81). F, Protein levels and quantitation results of SWAP70, ANP, BNP, MYH7 in NRVMs with ph PE (50 μmol/L) treatment at indicated time points (n=5 independent experiments). G, Relative mRNA levels of Swap70 in NRVMs with PE (50 μmol/L) treatment at indicated time points (n=5 independent experiments) (NS: left, P=1.00; middle, P=1.00; right, P=0.93). Data are shown as mean±SD. One-way ANOVA with Bonferroni’s post hoc analysis (A through C, E, G, MYH7 in F) or with Tamhane T2 post hoc test (MYH7 in A, D, F) was used. *P<0.05; **P<0.01 for indicated group vs control group. NS (no significance for indicated group vs control group. ANP indicates atrial natriuretic peptide; BNP, B-type natriuretic peptide; DCM, dilated cardiomyopathy; HCM, hypertrophic cardiomyopathy; MYH7/β-MHC, β-myosin-heavy-chain; NRVMs, neonatal rat ventricular myocytes; NS, no significance; PE, phenylephrine; SWAP70, switch-associated protein 70; and TAC, transverse aortic constriction.

Article Snippet: Manufacturer SWAP70 A14857 Abclonal SAPK/JNK 9252 CST Phospho-SAPK/JNK(Thr183/Tyr185) 4668 CST Phospho-p38 MAPK (Thr180/Tyr182) 4511 CST p38 MAPK 8690 CST Phospho-TAK1 (Thr184/187) 4508S CST TAK1 5206 CST ANP 27426-1-AP proteintech BNP A2179 Abclonal MYH7 22280-1-AP proteintech α-actinin 3134 CST Lamp1 1D4B DSHB Lamp1 9091 CST GRN A5773 Abclonal AKT A17909 Abclonal Phospho-AKT 4060 CST IKKβ A0714 Abclonal Phospho- IKKα/β 2694 CST Flag M185-3LL MBL Flag 20543-AP proteintech HA M180-3 MBL HA 3724S CST GAPDH 60004-1-Ig proteintech Goat anti-mouse IgG (H+L) 115-035-003 Jackson Goat anti-rabbit IgG (H+L) 111-035-003 Jackson D ow nloaded from http://ahajournals.org by on January 19, 2025 568 Goat anti-rabbit IgG (H+L) A11036 invitrogen 488 Goat anti-mouse IgG (H+L) A11029 invitrogen 568 Goat anti-mouse IgG (H+L) A11031 invitrogen 488 Goat anti-rabbit IgG (H+L) A11034 invitrogen D ow nloaded from http://ahajournals.org by on January 19, 2025 Table S2.

Techniques: Western Blot, Quantitation Assay, Expressing, Immunohistochemical staining, Staining, Control

Journal: Journal of the American Heart Association

Article Title: SWAP70 Overexpression Protects Against Pathological Cardiac Hypertrophy in a TAK1‐Dependent Manner

doi: 10.1161/jaha.122.028628

Figure Lengend Snippet: Figure 4. Swap70 overexpression suppresses the progression of cardiomyocyte hypertrophy. A, Immunoblotting (left) and quantitation (right) of SWAP70, ANP, BNP, and MYH7 protein levels in NRVMs infected with Ad-GFP- sh-Swap70 or Ad-sh-GFP in the presence or absence of PE (50 μmol/L) for 48 hours (n=5 independent experiments). B, Relative mRNA levels of Swap70 and hypertrophic marker genes (Anp, Bnp, and Myh7) in NRVMs from indicated groups (n=5 independent experiments). C, Representative immunofluorescence images (upper) of NRVMs infected with Ad-GFP-sh-Swap70 or Ad-sh-GFP adenovirus in the presence or absence of PE (50 μmol/L) for 24 hours and stained with α-actinin. Scale bar, 50 μm. Quantitative results (below) of the cardiomyocyte surface area from indicated groups (n=5 independent experiments). D, Immunoblotting (left) and quantitation (right) of SWAP70, ANP, BNP, MYH7 protein levels in NRVMs infected with Ad-GFP-Flag-Swap70 or Ad-GFP in the presence or absence of PE (50 μmol/L) for 48 hours (n=5 independent experiments). E, Relative mRNA levels of Swap70 and hypertrophic marker genes (Anp, Bnp, and Myh7) in NRVMs from indicated groups (n=5 independent experiments). F, Representative immunofluorescence images (upper) of NRVMs infected with Ad-GFP-Flag-Swap70 or Ad-GFP adenovirus in the presence or absence of PE (50 μmol/L) for 24 hours and stained with α-actinin. Scale bar, 50 μm. Quantitative results (below) of the cell surface area from indicated groups (n=5 independent experiments). Data are shown as mean±SD. Student t-test (A, D) and 1-way ANOVA with Bonferroni’s post hoc analysis (B, E, F) or with Tamhane T2 post hoc test (mRNA level of Myh7 in B, C) were used. *P<0.05; **P<0.01 for GFP PE group vs GFP PBS group. #P<0.05; ##P<0.01 for Swap70 overexpression/knockdown PE group vs GFP PE group. ANP indicates atrial natriuretic peptide; BNP, B-type natriuretic peptide; MYH7, β-myosin-heavy-chain; NRVMs, neonatal rat ventricular myocytes; PE, phenylephrine; and SWAP70, switch-associated protein 70.

Article Snippet: Manufacturer SWAP70 A14857 Abclonal SAPK/JNK 9252 CST Phospho-SAPK/JNK(Thr183/Tyr185) 4668 CST Phospho-p38 MAPK (Thr180/Tyr182) 4511 CST p38 MAPK 8690 CST Phospho-TAK1 (Thr184/187) 4508S CST TAK1 5206 CST ANP 27426-1-AP proteintech BNP A2179 Abclonal MYH7 22280-1-AP proteintech α-actinin 3134 CST Lamp1 1D4B DSHB Lamp1 9091 CST GRN A5773 Abclonal AKT A17909 Abclonal Phospho-AKT 4060 CST IKKβ A0714 Abclonal Phospho- IKKα/β 2694 CST Flag M185-3LL MBL Flag 20543-AP proteintech HA M180-3 MBL HA 3724S CST GAPDH 60004-1-Ig proteintech Goat anti-mouse IgG (H+L) 115-035-003 Jackson Goat anti-rabbit IgG (H+L) 111-035-003 Jackson D ow nloaded from http://ahajournals.org by on January 19, 2025 568 Goat anti-rabbit IgG (H+L) A11036 invitrogen 488 Goat anti-mouse IgG (H+L) A11029 invitrogen 568 Goat anti-mouse IgG (H+L) A11031 invitrogen 488 Goat anti-rabbit IgG (H+L) A11034 invitrogen D ow nloaded from http://ahajournals.org by on January 19, 2025 Table S2.

Techniques: Over Expression, Western Blot, Quantitation Assay, Infection, Marker, Immunofluorescence, Staining, Knockdown

Journal: Journal of the American Heart Association

Article Title: SWAP70 Overexpression Protects Against Pathological Cardiac Hypertrophy in a TAK1‐Dependent Manner

doi: 10.1161/jaha.122.028628

Figure Lengend Snippet: Figure 6. SWAP70 regulates cardiomyocyte hypertrophy by interacting with TAK1. A, Representative immunofluorescence images of the colocalization of HA-tagged SWAP70 and endogenous TAK1 in NRVMs infected with Ad-HA-Swap70 adenovirus in the presence of PE (50 μmol/L) for 24 hours. Scale bar, 25 μm. B, CO-IP assays displayed the interaction between endogenous TAK1 and Flag-tagged SWAP70 in NRVMs. C, CO-IP assays displayed the interaction between endogenous TAK1 and Flag-tagged SWAP70 (193–585) in NRVMs. D, Immunoblotting analysis of phosphorylation and total TAK1, P38, JNK1/2 proteins in NRVMs infected with indicated adenoviruses and stimulated with PE (50 μmol/L) for 48 hours, hypertrophic marker proteins (ANP, BNP, and MYH7) were used to assess the severity of the hypertrophy (n=5 independent experiments). E, Relative mRNA levels of hypertrophic marker genes (Anp, Bnp, and Myh7) in NRVMs from indicated groups (n=5 independent experiments) (NS: high, P=0.25; middle, P=0.08; low, P=0.65). F, Representative immunofluorescence images (left) of NRVMs infected with indicated adenovirus in the presence of PE (50 μmol/L) for 24 hours and stained with α-actinin. Scale bar, 50 μm. Quantitative results (right) of the cell surface area from indicated groups (n=5 independent experiments) (NS: P=0.15). Data are shown as mean±SD. One-way ANOVA with Bonferroni’s post hoc analysis (E, F) or with Tamhane T2 post hoc test (mRNA levels of Myh7 in E) were used. ##P<0.01 for indicated group vs control group. NS for indicated group vs control group. ANP indicates atrial natriuretic peptide; BNP, B-type natriuretic peptide; CO-IP, coimmunoprecipitation; JNK, c-Jun N-terminal kinase; MHY7, β-myosin-heavy-chain; NRVMs, neonatal rat ventricular myocytes; NS, no significance; PE, phenylephrine; SWAP70, switch-associated protein 70; and TAK1, transforming growth factor β-activated kinase 1.

Article Snippet: Manufacturer SWAP70 A14857 Abclonal SAPK/JNK 9252 CST Phospho-SAPK/JNK(Thr183/Tyr185) 4668 CST Phospho-p38 MAPK (Thr180/Tyr182) 4511 CST p38 MAPK 8690 CST Phospho-TAK1 (Thr184/187) 4508S CST TAK1 5206 CST ANP 27426-1-AP proteintech BNP A2179 Abclonal MYH7 22280-1-AP proteintech α-actinin 3134 CST Lamp1 1D4B DSHB Lamp1 9091 CST GRN A5773 Abclonal AKT A17909 Abclonal Phospho-AKT 4060 CST IKKβ A0714 Abclonal Phospho- IKKα/β 2694 CST Flag M185-3LL MBL Flag 20543-AP proteintech HA M180-3 MBL HA 3724S CST GAPDH 60004-1-Ig proteintech Goat anti-mouse IgG (H+L) 115-035-003 Jackson Goat anti-rabbit IgG (H+L) 111-035-003 Jackson D ow nloaded from http://ahajournals.org by on January 19, 2025 568 Goat anti-rabbit IgG (H+L) A11036 invitrogen 488 Goat anti-mouse IgG (H+L) A11029 invitrogen 568 Goat anti-mouse IgG (H+L) A11031 invitrogen 488 Goat anti-rabbit IgG (H+L) A11034 invitrogen D ow nloaded from http://ahajournals.org by on January 19, 2025 Table S2.

Techniques: Immunofluorescence, Infection, Co-Immunoprecipitation Assay, Western Blot, Phospho-proteomics, Marker, Staining, Control

Journal: Journal of the American Heart Association

Article Title: SWAP70 Overexpression Protects Against Pathological Cardiac Hypertrophy in a TAK1‐Dependent Manner

doi: 10.1161/jaha.122.028628

Figure Lengend Snippet: Figure 8. Schematic showing possible mechanisms underlying normal or hypertrophic environment associated with SWAP70. Under normal conditions, SWAP70 is degraded by lysosomes mediated by GRN to maintain the balance of SWAP70 protein level, while hypertrophic stress reduces the binding of SWAP70 to GRN and thus decreases the lysosomal degradation of SWAP70, leads to the increased SWAP70 protein level, decreases the interaction between TAK1 and TAB1, inhibits the phosphorylation of TAK1- JNK 1/2-P38 signaling pathways, and finally suppresses cardiac hypertrophy. The phosphorylation inhibition of TAK1 by SWAP70 is dependent on its N-terminal interaction with TAK1. GRN indicates granulin precursor; JNK, c-Jun N-terminal kinase; SWAP70, switch- associated protein 70; TAB1, transforming growth factor β-activated kinase 1 binding protein 1; and TAK1, transforming growth factor β-activated kinase 1.

Article Snippet: Manufacturer SWAP70 A14857 Abclonal SAPK/JNK 9252 CST Phospho-SAPK/JNK(Thr183/Tyr185) 4668 CST Phospho-p38 MAPK (Thr180/Tyr182) 4511 CST p38 MAPK 8690 CST Phospho-TAK1 (Thr184/187) 4508S CST TAK1 5206 CST ANP 27426-1-AP proteintech BNP A2179 Abclonal MYH7 22280-1-AP proteintech α-actinin 3134 CST Lamp1 1D4B DSHB Lamp1 9091 CST GRN A5773 Abclonal AKT A17909 Abclonal Phospho-AKT 4060 CST IKKβ A0714 Abclonal Phospho- IKKα/β 2694 CST Flag M185-3LL MBL Flag 20543-AP proteintech HA M180-3 MBL HA 3724S CST GAPDH 60004-1-Ig proteintech Goat anti-mouse IgG (H+L) 115-035-003 Jackson Goat anti-rabbit IgG (H+L) 111-035-003 Jackson D ow nloaded from http://ahajournals.org by on January 19, 2025 568 Goat anti-rabbit IgG (H+L) A11036 invitrogen 488 Goat anti-mouse IgG (H+L) A11029 invitrogen 568 Goat anti-mouse IgG (H+L) A11031 invitrogen 488 Goat anti-rabbit IgG (H+L) A11034 invitrogen D ow nloaded from http://ahajournals.org by on January 19, 2025 Table S2.

Techniques: Binding Assay, Phospho-proteomics, Protein-Protein interactions, Inhibition

Journal: Foods

Article Title: Role and Mechanism of Sialic Acid in Alleviating Acute Lung Injury through In Vivo and In Vitro Models

doi: 10.3390/foods13182984

Figure Lengend Snippet: Effects of SA supplementation on the JNK/p38/PPAR-γ-NF-κB pathway and the expressions of the oxidation-related genes of the lung tissues. The protein expressions of JNK, p38, NF-κB, and PPAR-γ were tested by western blot analysis ( A , B ). Effects of SA supplementation on the protein levels of oxidation-related genes (HO-1 and NQO1) ( C , D ). CON: control; LPS: lipopolysaccharide; LPS + 50 SA: LPS + 50 mg/kg/day SA; LPS + 100 SA: LPS + 100 mg/kg/day SA; SA: sialic acid. The data are expressed as mean ± SD. CON group vs. LPS group or LPS group vs. LPS + 50 SA group and LPS + 100 SA group. * p < 0.05. ** p < 0.01. # p > 0.05.

Article Snippet: Polyclonal antibody tumor necrosis factor-α (TNF-α, CAT. #6945S), interleukin-6 (IL-6, CAT. #12912S), interleukin-1β (IL-1β, CAT. #12242S), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 (CAT. #12629S), activator protein 1 (AP-1, CAT. #9165S), and mitogen-activated protein kinase (MAPK) family antibody (JNK, CAT. #9252S; p-JNK, CAT. #4668S; p38, CAT. #9212S; p-p38, CAT. #4511S) were provided by Cell Signaling Technology (Danver, MA, USA).

Techniques: Western Blot, Control

Journal: Foods

Article Title: Role and Mechanism of Sialic Acid in Alleviating Acute Lung Injury through In Vivo and In Vitro Models

doi: 10.3390/foods13182984

Figure Lengend Snippet: Effects of SA supplementation on the p38/JNK and Nrf2 pathways in LPS-induced HUVEC cells. HUVEC cells were pretreated with different dosages of SA (25, 50, and 100 μg/mL) for 2 h and then treated with LPS for 10–12 h. The total protein was obtained from the cells, and the protein expressions of p38, p-p38, JNK, and p-JNK were determined by western blot analysis ( A , B ). The effects of SA supplementation on the Nrf2 pathway ( C , D ). LPS: lipopolysaccharide; SA: sialic acid. Data are presented as the mean ± SD of three independent experiments. CON group vs. LPS group or LPS group vs. 25, 50, and 100 μg/mL SA group. * p < 0.05, ** p < 0.01. # p > 0.05.

Article Snippet: Polyclonal antibody tumor necrosis factor-α (TNF-α, CAT. #6945S), interleukin-6 (IL-6, CAT. #12912S), interleukin-1β (IL-1β, CAT. #12242S), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 (CAT. #12629S), activator protein 1 (AP-1, CAT. #9165S), and mitogen-activated protein kinase (MAPK) family antibody (JNK, CAT. #9252S; p-JNK, CAT. #4668S; p38, CAT. #9212S; p-p38, CAT. #4511S) were provided by Cell Signaling Technology (Danver, MA, USA).

Techniques: Western Blot

Journal: Foods

Article Title: Role and Mechanism of Sialic Acid in Alleviating Acute Lung Injury through In Vivo and In Vitro Models

doi: 10.3390/foods13182984

Figure Lengend Snippet: Schematic representation of the protective effect and possible molecular mechanisms of SA on ALI. SA exerted a protective effect against LPS-induced ALI via ameliorating inflammation and oxidative damage, which worked mainly through regulation of the JNK/p38-NF-κB/AP-1 and Nrf2 signaling pathways. ALI: acute lung injury; HUVEC: human umbilical vein endothelial cells; LPS: lipopolysaccharide; SA: sialic acid.

Article Snippet: Polyclonal antibody tumor necrosis factor-α (TNF-α, CAT. #6945S), interleukin-6 (IL-6, CAT. #12912S), interleukin-1β (IL-1β, CAT. #12242S), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) p65 (CAT. #12629S), activator protein 1 (AP-1, CAT. #9165S), and mitogen-activated protein kinase (MAPK) family antibody (JNK, CAT. #9252S; p-JNK, CAT. #4668S; p38, CAT. #9212S; p-p38, CAT. #4511S) were provided by Cell Signaling Technology (Danver, MA, USA).

Techniques: