Journal: Pathophysiology

Article Title: Altered Production and Cellular Levels of Hydrogen Sulfide (H2S) in Placental Trophoblasts from Pregnancies Affected by Pre-Eclampsia

doi: 10.3390/pathophysiology32010010

Figure Lengend Snippet: Figure 2. Expression of CBS, CSE, and 3-MST, as determined by immunostaining in placental villous tis- sue. (A) Representative CBS, CSE, and 3-MST expression, detected by immunostaining from normal and pre-eclamptic cases. In the normal group, both CBS and CSE were detected in the syncytiotrophoblast layer (double arrow). CBS, but not CSE, was also detected in the villous core fetal vessel endothelium (solid arrowhead). Expression of CBS and CSE were markedly reduced in syncytiotrophoblasts in pre-eclamptic cases (Figure 2A(b,d)) compared to controls. Expression of 3-MST was undetectable in placental specimens from both normal and pre-eclamptic cases (Figure 2A(e,f)). Bar = 50 µm. (B) Repre- sentative CBS, CSE, and 3-MST expression in the first, second, and third trimester placental specimens. Both CBS and CSE are strongly expressed in cyto- and syncytio-trophoblasts in the first, second, and third trimester placentas. CBS: a, b, and c; CSE: d, e, and f, respectively. Expression of 3-MST was detected in the first (g) and the second (h) trimester placentas, but not in the third/term (i) placentas. Bar = 50 µm.

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM, D5523), Percoll (P1644), Hanks balanced salt solution (HBSS, H4891), Monobrombimane (MBB, B4380), and antibody against β-actin (A2066) were purchased from Sigma-Aldrich (St. Louis, MO, USA); Trypsin (LS003703) and Deoxyribonuclease (LS002139) were purchased from Worthington Biochemical Corporation (Lakewood, NJ, USA); fetal bovine serum (FBS) was from Atlantic Biologicals (Flowery Branch, GA, USA); Antibodies against CBS (12217-1-AP) and CSE (14787-1-AP) were purchased from Proteintech (Rosemont, IL, USA); and antibody against 3-MST (AB224043) was from Abcam (Waltham, MA, USA).

Techniques: Expressing, Immunostaining

Journal: Pathophysiology

Article Title: Altered Production and Cellular Levels of Hydrogen Sulfide (H2S) in Placental Trophoblasts from Pregnancies Affected by Pre-Eclampsia

doi: 10.3390/pathophysiology32010010

Figure Lengend Snippet: Figure 3. Trophoblast CBS and CSE protein expression by Western blot. Consistent with immunos- taining results, the levels of CBS and CSE protein expression were also significantly reduced in trophoblasts from pre-eclamptic (PE, n = 6) compared to those from normal (n = 7) cases. The bar graphs show relative protein expression of CBS and CSE after normalization with β-actin expression for each sample, * p < 0.05 and ** p < 0.01: PE vs. normal, respectively.

Article Snippet: Dulbecco’s modified Eagle’s medium (DMEM, D5523), Percoll (P1644), Hanks balanced salt solution (HBSS, H4891), Monobrombimane (MBB, B4380), and antibody against β-actin (A2066) were purchased from Sigma-Aldrich (St. Louis, MO, USA); Trypsin (LS003703) and Deoxyribonuclease (LS002139) were purchased from Worthington Biochemical Corporation (Lakewood, NJ, USA); fetal bovine serum (FBS) was from Atlantic Biologicals (Flowery Branch, GA, USA); Antibodies against CBS (12217-1-AP) and CSE (14787-1-AP) were purchased from Proteintech (Rosemont, IL, USA); and antibody against 3-MST (AB224043) was from Abcam (Waltham, MA, USA).

Techniques: Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Hydrogen Sulfide Reduces Myeloid-Derived Suppressor Cell-Mediated Inflammatory Response in a Model of Helicobacter hepaticus -Induced Colitis

doi: 10.3389/fimmu.2018.00499

Figure Lengend Snippet: H 2 S production is significantly reduced during Helicobacter hepaticus ( Hh )-induced colitis. 129SvEv Rag2 −/− mice were infected with Hh and sacrificed 6 weeks post-infection. (A) Representative western blot and (B) relative CSE and CBS mRNA expression performed on colon samples from uninfected and Hh -infected mice. A significant reduction in the expression of both CBS protein and mRNA was observed in Hh -infected mice. (C) H 2 S synthesis by the colon of healthy mice was measured in colon homogenates, in the absence or presence of CBS inhibitor [(CHH), 3 mmol/L] or CSE inhibitor (PAG, 10 mmol/L). The substrate for H 2 S synthesis ( l -cys) was present at 4 mmol/L. Only CHH significantly reduced H 2 S synthesis in tissue from healthy controls (*** P < 0.001 vs. l -cys alone). (D) H 2 S levels measured in the colon of Hh -infected mice 3 or 6 weeks post-infection resulted significantly reduced as compared to uninfected mice. Data are shown as mean ± SEM ( n = 5 per group) (** P < 0.01, *** P < 0.001 vs. uninfected).

Article Snippet: Then, the filters were incubated with the following primary antibodies: CSE (1:500 dilution, Proteintech), CBS (1:500, Novus Biological), and GAPDH (1:1,000 dilution, Santa Cruz Biotechnology) overnight at 4°C.

Techniques: Infection, Western Blot, Expressing

Journal: EMBO molecular medicine

Article Title: Mitochondrial MsrB2 serves as a switch and transducer for mitophagy.

doi: 10.15252/emmm.201910409

Figure Lengend Snippet: Figure 1. Identification of MsrB2 as LC3 interaction protein in human platelets.

Article Snippet: Healthy/DM platelet lysates and cell lysates (after transient transfection) were mixed with the specific target antibody (1 lg of LC3 antirabbit antibody (Abcam) and 2 lg of Parkin anti-goat antibody (Abcam) or 1.5 lg of Parkin anti-rabbit antibody (Abcam) for Parkin IP; 1.5 lg of MsrB2 anti-rabbit antibody (laboratory made) for MsrB2 IP; and 1 lg of GFP anti-rabbit antibody (Abcam, USA) or GFP-Trap bead (Chromotek, USA), and the same species IgG control with HC) and incubated overnight at 4°C.

Techniques:

Journal: EMBO molecular medicine

Article Title: Mitochondrial MsrB2 serves as a switch and transducer for mitophagy.

doi: 10.15252/emmm.201910409

Figure Lengend Snippet: Figure 2. MsrB2 is needed for mitophagy.

Article Snippet: Healthy/DM platelet lysates and cell lysates (after transient transfection) were mixed with the specific target antibody (1 lg of LC3 antirabbit antibody (Abcam) and 2 lg of Parkin anti-goat antibody (Abcam) or 1.5 lg of Parkin anti-rabbit antibody (Abcam) for Parkin IP; 1.5 lg of MsrB2 anti-rabbit antibody (laboratory made) for MsrB2 IP; and 1 lg of GFP anti-rabbit antibody (Abcam, USA) or GFP-Trap bead (Chromotek, USA), and the same species IgG control with HC) and incubated overnight at 4°C.

Techniques:

Journal: EMBO molecular medicine

Article Title: Mitochondrial MsrB2 serves as a switch and transducer for mitophagy.

doi: 10.15252/emmm.201910409

Figure Lengend Snippet: Figure 5. Methionine sulfoxidation (MetO) and methionine sulfide reductase B2 (MsrB2) are increased, along with Parkin, in DM platelets.

Article Snippet: Healthy/DM platelet lysates and cell lysates (after transient transfection) were mixed with the specific target antibody (1 lg of LC3 antirabbit antibody (Abcam) and 2 lg of Parkin anti-goat antibody (Abcam) or 1.5 lg of Parkin anti-rabbit antibody (Abcam) for Parkin IP; 1.5 lg of MsrB2 anti-rabbit antibody (laboratory made) for MsrB2 IP; and 1 lg of GFP anti-rabbit antibody (Abcam, USA) or GFP-Trap bead (Chromotek, USA), and the same species IgG control with HC) and incubated overnight at 4°C.

Techniques:

Journal: EMBO molecular medicine

Article Title: Mitochondrial MsrB2 serves as a switch and transducer for mitophagy.

doi: 10.15252/emmm.201910409

Figure Lengend Snippet: Figure 6. MsrB2 is ubiquitinated by Parkin.

Article Snippet: Healthy/DM platelet lysates and cell lysates (after transient transfection) were mixed with the specific target antibody (1 lg of LC3 antirabbit antibody (Abcam) and 2 lg of Parkin anti-goat antibody (Abcam) or 1.5 lg of Parkin anti-rabbit antibody (Abcam) for Parkin IP; 1.5 lg of MsrB2 anti-rabbit antibody (laboratory made) for MsrB2 IP; and 1 lg of GFP anti-rabbit antibody (Abcam, USA) or GFP-Trap bead (Chromotek, USA), and the same species IgG control with HC) and incubated overnight at 4°C.

Techniques:

Journal: EMBO molecular medicine

Article Title: Mitochondrial MsrB2 serves as a switch and transducer for mitophagy.

doi: 10.15252/emmm.201910409

Figure Lengend Snippet: Figure 7. MsrB2 is released by mitochondrial rupture.

Article Snippet: Healthy/DM platelet lysates and cell lysates (after transient transfection) were mixed with the specific target antibody (1 lg of LC3 antirabbit antibody (Abcam) and 2 lg of Parkin anti-goat antibody (Abcam) or 1.5 lg of Parkin anti-rabbit antibody (Abcam) for Parkin IP; 1.5 lg of MsrB2 anti-rabbit antibody (laboratory made) for MsrB2 IP; and 1 lg of GFP anti-rabbit antibody (Abcam, USA) or GFP-Trap bead (Chromotek, USA), and the same species IgG control with HC) and incubated overnight at 4°C.

Techniques: