anti cytoc  (Cell Signaling Technology Inc)

Journal: Pharmaceuticals

Article Title: Comparison of Quercetin and Isoquercitrin’s Anti-Heart Failure Activity via MAPK Inflammatory Pathway and Caspase Apoptosis Pathway

doi: 10.3390/ph18101447

Figure Lengend Snippet: WB analysis of inflammatory and apoptotic proteins in cardiomyocytes treated with IQ and Que in vitro. ( A ) IQ and Que regulate Ang II induced expression of apoptosis related proteins (Caspase-3/Bax/CytoC/Bcl-2) in H9c2 cells. Grayscale quantification of apoptotic markers (Bax/Bcl-2/Caspase-3/CytoC). ( B ) IQ and Que modulate Ang II induced expression of JNK/ERK/P38 and their phosphorylated forms in H9c2 cells. Quantification of inflammatory protein expression by densitometric analysis. Data are presented as mean ± SD. (n = 3 data were presented as mean ± SEM, # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, vs. Ang II, ** p < 0.01, IQ vs. Que, ns, p > 0.05, IQ vs. Que).

Article Snippet: Bicinchoninic acid (BCA) protein detection kit (item number: PC0020), Hoechst 33342 (item number: C0030), PI (item number: C0080), (Solarbio, Beijing, China), Ang II (item number: S25704), IQ (item number: S33219 ), Que (item number: S33752), CCK-8 (item number: R22305 ), DCFH-DA (item number: R22380 ), RIPA lysis buffer (item number: R32714 ), (Yuanye, Shanghai, China), sodium dodecyl sulfate polyacrylamide gel electrophoresis preparation kit (SDS-PAGE) (item number: BL522A), electrochemiluminescence (ECL) substrate kit (item number: BL520A), (Biosharp, Beijing, China), p-ERK (item number: 28733-1-AP), ERK (item number: 11257-1-AP), p-JNK (item number: 80024-1-RR), JNK (item number: 51153-1-AP), p-P38 (item number: 28796-1-AP) and P38 (item number: 14064-1-AP), Caspase-3 (item number: 19677-1-AP), Bax (item number: 50599-2-Ig), Bcl-2 (item number: 12789-1-AP), CytoC (item number: 10993-1-AP), GADPH (item number: 10494-1-AP), β-actin (item number: 20536-1-AP), HRP-conjugated Goat Anti-Rabbit IgG(H+L) (item number: SA00001-2), (Proteintech, Chicago, IL, USA), polyvinylidene fluoride (PVDF) membrane (item number: IPVH00010), (Merck, Darmstadt, Germany), fluorescence microscope (Model No. 10119978), (Nikon, Tokyo, Japan), CO 2 incubator (Device Model: BC-J250), (Thermo, Waltham, MA, USA), electrophoresis apparatus (Serial No. 041BR174116), (BIO-RAD, Hercules, CA, USA).

Techniques: In Vitro, Expressing

Journal: Pharmaceuticals

Article Title: Comparison of Quercetin and Isoquercitrin’s Anti-Heart Failure Activity via MAPK Inflammatory Pathway and Caspase Apoptosis Pathway

doi: 10.3390/ph18101447

Figure Lengend Snippet: Effects of IQ and Que on apoptotic pathways in Ang II-induced myocardial injured mice. ( A ) IQ and Que regulate Ang II induced apoptosis related proteins (Caspase-3/Bax/CytoC/Bcl-2) in C57BL/6J mice. In vivo quantification of apoptotic protein expression by densitometric analysis. ( B ) Effects of IQ and Que on Ang II-induced JNK/ERK/P38 phosphorylation in C57BL/6J mice. In vivo analysis of inflammatory protein expression. (n = 3 data were presented as mean ± SEM, # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001, vs. Ang II, ns, p > 0.05, IQ vs. Que).

Article Snippet: Bicinchoninic acid (BCA) protein detection kit (item number: PC0020), Hoechst 33342 (item number: C0030), PI (item number: C0080), (Solarbio, Beijing, China), Ang II (item number: S25704), IQ (item number: S33219 ), Que (item number: S33752), CCK-8 (item number: R22305 ), DCFH-DA (item number: R22380 ), RIPA lysis buffer (item number: R32714 ), (Yuanye, Shanghai, China), sodium dodecyl sulfate polyacrylamide gel electrophoresis preparation kit (SDS-PAGE) (item number: BL522A), electrochemiluminescence (ECL) substrate kit (item number: BL520A), (Biosharp, Beijing, China), p-ERK (item number: 28733-1-AP), ERK (item number: 11257-1-AP), p-JNK (item number: 80024-1-RR), JNK (item number: 51153-1-AP), p-P38 (item number: 28796-1-AP) and P38 (item number: 14064-1-AP), Caspase-3 (item number: 19677-1-AP), Bax (item number: 50599-2-Ig), Bcl-2 (item number: 12789-1-AP), CytoC (item number: 10993-1-AP), GADPH (item number: 10494-1-AP), β-actin (item number: 20536-1-AP), HRP-conjugated Goat Anti-Rabbit IgG(H+L) (item number: SA00001-2), (Proteintech, Chicago, IL, USA), polyvinylidene fluoride (PVDF) membrane (item number: IPVH00010), (Merck, Darmstadt, Germany), fluorescence microscope (Model No. 10119978), (Nikon, Tokyo, Japan), CO 2 incubator (Device Model: BC-J250), (Thermo, Waltham, MA, USA), electrophoresis apparatus (Serial No. 041BR174116), (BIO-RAD, Hercules, CA, USA).

Techniques: In Vivo, Expressing, Phospho-proteomics

Journal: Pharmaceuticals

Article Title: Comparison of Quercetin and Isoquercitrin’s Anti-Heart Failure Activity via MAPK Inflammatory Pathway and Caspase Apoptosis Pathway

doi: 10.3390/ph18101447

Figure Lengend Snippet: Molecular docking analysis of compounds with core targets. ( A ) IQ with the Caspase-3; ( B ) IQ with the Bcl-2; ( C ) IQ with the Bax; ( D ) IQ with CytoC; ( E ) IQ with JNK; ( F ) IQ with ERK; ( G ) IQ with P38.

Article Snippet: Bicinchoninic acid (BCA) protein detection kit (item number: PC0020), Hoechst 33342 (item number: C0030), PI (item number: C0080), (Solarbio, Beijing, China), Ang II (item number: S25704), IQ (item number: S33219 ), Que (item number: S33752), CCK-8 (item number: R22305 ), DCFH-DA (item number: R22380 ), RIPA lysis buffer (item number: R32714 ), (Yuanye, Shanghai, China), sodium dodecyl sulfate polyacrylamide gel electrophoresis preparation kit (SDS-PAGE) (item number: BL522A), electrochemiluminescence (ECL) substrate kit (item number: BL520A), (Biosharp, Beijing, China), p-ERK (item number: 28733-1-AP), ERK (item number: 11257-1-AP), p-JNK (item number: 80024-1-RR), JNK (item number: 51153-1-AP), p-P38 (item number: 28796-1-AP) and P38 (item number: 14064-1-AP), Caspase-3 (item number: 19677-1-AP), Bax (item number: 50599-2-Ig), Bcl-2 (item number: 12789-1-AP), CytoC (item number: 10993-1-AP), GADPH (item number: 10494-1-AP), β-actin (item number: 20536-1-AP), HRP-conjugated Goat Anti-Rabbit IgG(H+L) (item number: SA00001-2), (Proteintech, Chicago, IL, USA), polyvinylidene fluoride (PVDF) membrane (item number: IPVH00010), (Merck, Darmstadt, Germany), fluorescence microscope (Model No. 10119978), (Nikon, Tokyo, Japan), CO 2 incubator (Device Model: BC-J250), (Thermo, Waltham, MA, USA), electrophoresis apparatus (Serial No. 041BR174116), (BIO-RAD, Hercules, CA, USA).

Techniques:

Journal: Pharmaceuticals

Article Title: Comparison of Quercetin and Isoquercitrin’s Anti-Heart Failure Activity via MAPK Inflammatory Pathway and Caspase Apoptosis Pathway

doi: 10.3390/ph18101447

Figure Lengend Snippet: Molecular docking analysis of compounds with core targets. ( A ) Que with the Caspase-3; ( B ) Que with the Bcl-2; ( C ) Que with the Bax; ( D ) Que with CytoC; ( E ) Que with JNK; ( F ) Que with ERK; ( G ) Que with P38.

Article Snippet: Bicinchoninic acid (BCA) protein detection kit (item number: PC0020), Hoechst 33342 (item number: C0030), PI (item number: C0080), (Solarbio, Beijing, China), Ang II (item number: S25704), IQ (item number: S33219 ), Que (item number: S33752), CCK-8 (item number: R22305 ), DCFH-DA (item number: R22380 ), RIPA lysis buffer (item number: R32714 ), (Yuanye, Shanghai, China), sodium dodecyl sulfate polyacrylamide gel electrophoresis preparation kit (SDS-PAGE) (item number: BL522A), electrochemiluminescence (ECL) substrate kit (item number: BL520A), (Biosharp, Beijing, China), p-ERK (item number: 28733-1-AP), ERK (item number: 11257-1-AP), p-JNK (item number: 80024-1-RR), JNK (item number: 51153-1-AP), p-P38 (item number: 28796-1-AP) and P38 (item number: 14064-1-AP), Caspase-3 (item number: 19677-1-AP), Bax (item number: 50599-2-Ig), Bcl-2 (item number: 12789-1-AP), CytoC (item number: 10993-1-AP), GADPH (item number: 10494-1-AP), β-actin (item number: 20536-1-AP), HRP-conjugated Goat Anti-Rabbit IgG(H+L) (item number: SA00001-2), (Proteintech, Chicago, IL, USA), polyvinylidene fluoride (PVDF) membrane (item number: IPVH00010), (Merck, Darmstadt, Germany), fluorescence microscope (Model No. 10119978), (Nikon, Tokyo, Japan), CO 2 incubator (Device Model: BC-J250), (Thermo, Waltham, MA, USA), electrophoresis apparatus (Serial No. 041BR174116), (BIO-RAD, Hercules, CA, USA).

Techniques:

Journal: Nutrients

Article Title: Histidine Triad Nucleotide-Binding Protein 1 Improves Critical Limb Ischemia by Regulating Mitochondrial Homeostasis

doi: 10.3390/nu15234859

Figure Lengend Snippet: Silencing of Hint1 damages mitochondrial function in EC. ( A ) An analysis of O 2 consumption in HUVECs using different inhibitors. The ECs were treated with HG (25 mM) or vehicle after transfection with the siHint1 for 24 h. Graphs present the basal OCR ( B ), maximal OCR ( C ), ATP production ( D ), and spare respiratory capacity OCR ( E ) in ECs; n = 3 per group, two-way ANOVA with Bonferroni multiple comparison test. ** p < 0.01. ( F , G ) MitoTracker staining was used to detect mitochondrial membrane potential (Δψm); Scar bar = 100 μm, n = 5 per group, two-way ANOVA with Bonferroni multiple comparison test. * p < 0.05, *** p < 0.001. ( H , I ) mitoSOX fluorescence staining was used to detect mitochondrial reactive oxygen species (mROS); Scar bar = 50 μm, n = 5 per group, two-way ANOVA with Bonferroni multiple comparison test. ** p < 0.01. ( J ) Representative Western blot analysis of SOD2, CATALASE, and CytoC in ECs are shown.

Article Snippet: The membranes were blocked using 5% ( w / v ) BSA in TBST and incubated with the respective primary antibodies: HINT1 (ab124912, Cambridge, MA, USA), SOD2 (PA5-30604, Invitrogen, Carlsbad, CA, USA), CATALASE (#8841, CST, Danvers, MA, USA), CytoC (#4272, CST, Danvers, MA, USA), and β-actin (#3700, CST, Danvers, MA, USA).

Techniques: Transfection, Comparison, Staining, Membrane, Fluorescence, Western Blot

Journal: Nutrients

Article Title: Histidine Triad Nucleotide-Binding Protein 1 Improves Critical Limb Ischemia by Regulating Mitochondrial Homeostasis

doi: 10.3390/nu15234859

Figure Lengend Snippet: Hint1 overexpression ameliorates mitochondrial dysfunction in ECs. ( A ) An analysis of O 2 consumption in HUVECs using different inhibitors. The ECs were treated with HG (25 mM) or vehicle after being infected with AAV-Hint1 for 48 h. Graphs present the basal OCR ( B ), maximal OCR ( C ), ATP production ( D ), and spare respiratory capacity OCR ( E ) in ECs; n = 3 per group, two-way ANOVA with Bonferroni multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001. ( F , G ) MitoTracker staining was used to detect mitochondrial membrane potential (Δψm); Scar bar = 100 μm, n = 5 per group, two-way ANOVA with Bonferroni multiple comparison test. * p < 0.05. ( H , I ) mitoSOX fluorescence staining was used to detect mitochondrial reactive oxygen species (mROS); Scar bar = 50 μm, n = 5 per group, two-way ANOVA with Bonferroni multiple comparison test. * p < 0.05. ( J ) Representative Western blot analysis of SOD2, CATALASE, and CytoC in ECs are shown.

Article Snippet: The membranes were blocked using 5% ( w / v ) BSA in TBST and incubated with the respective primary antibodies: HINT1 (ab124912, Cambridge, MA, USA), SOD2 (PA5-30604, Invitrogen, Carlsbad, CA, USA), CATALASE (#8841, CST, Danvers, MA, USA), CytoC (#4272, CST, Danvers, MA, USA), and β-actin (#3700, CST, Danvers, MA, USA).

Techniques: Over Expression, Infection, Comparison, Staining, Membrane, Fluorescence, Western Blot