rabbit polyclonal anti mat2a  (Proteintech)

Journal: Brain research bulletin

Article Title: MiR-26b-5p-modified hUB-MSCs derived exosomes attenuate early brain injury during subarachnoid hemorrhage via MAT2A-mediated the p38 MAPK/STAT3 signaling pathway.

doi: 10.1016/j.brainresbull.2021.07.014

Figure Lengend Snippet: Fig. 4. MAT2A was a direct target of miR- 26b-5p. A. StarBase3.0 (http://starbase.sysu.edu.cn/) was used to predicted the targeting sites of miR- 26b-5p and MAT2A. The wild-type and mutant sequences of MAT2A are shown in the figure. B. The relative luciferase activity was tested with wild-type and mutant-type of MAT2A, respec tively. C. The miR-26b-5p mimic or miR-26b-5p inhibitor was transfected into cells and the protein expression of MAT2A was analyzed by Western blotting. GAPDH was used as an in ternal reference. N = 6, *P < 0.01.

Article Snippet: Then, equal amounts of proteins (15 μg/lane) were separated on a 12 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenedifluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA).The membranes were blocked in 5% (w/v) nonfat dry milk in TBST (Tris-buffered saline-0.1 % Tween) at 25 ◦C for 3 h and then incubated with the following primary antibodies: rabbit polyclonal anti-GAPDH antibody (1:1000, Abcam, ab8245), rabbit monoclonal anti-CD63 antibody (1:1500, Abcam, ab217345), mouse monoclonal anti-CD81 antibody (1:1200, Abcam, ab79559), rabbit monoclonal anti-TSG101 antibody (1:2500, Abcam, ab125011), rabbit monoclonal anti-Alix antibody (1:2000, Abcam, ab186492), Rabbit polyclonal anti-MAT2A antibody (1:2500, Abcam, ab154343), Rabbit monoclonal anti-COX-2 antibody (1:2000, Abcam, ab15191), Rabbit monoclonal anti-MCP-1 antibody (1:1000, Abcam, ab214819), Rabbit monoclonal anti-iNOS antibody (1:800, Abcam, ab178945), rabbit monoclonal anti-STAT3 antibody (1:1500, Abcam, ab68153), rabbit monoclonal anti-AKT (phospho Y705) antibody (1:5000, Abcam, ab76315).

Techniques: Mutagenesis, Luciferase, Activity Assay, Transfection, Expressing, Western Blot

Journal: Brain research bulletin

Article Title: MiR-26b-5p-modified hUB-MSCs derived exosomes attenuate early brain injury during subarachnoid hemorrhage via MAT2A-mediated the p38 MAPK/STAT3 signaling pathway.

doi: 10.1016/j.brainresbull.2021.07.014

Figure Lengend Snippet: Fig. 5. Effect of regulating MAT2A expression on oxyHb-induced PC12 cells. PC12 cells were stimulated with 50 ng/mL oxyHb (cell density was 1 × 106) for 6 h. The cells were then incubated with exosomes at a concentration of 100 μg/mL, and the overexpression vector of MAT2A was transfected into the cells. A. The protein expression of MAT2A was analyzed by Western blotting. B. MTT assay was used to detected cell proliferation. C. Apoptosis of PC12 cells were detected by flow cytometry. D. Western botting was used to detected the protein expression of COX-2, MCP-1 and iNOS. GAPDH was used as an invariant internal control for calculating protein-fold changes. N = 6, *P < 0.01.

Article Snippet: Then, equal amounts of proteins (15 μg/lane) were separated on a 12 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenedifluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA).The membranes were blocked in 5% (w/v) nonfat dry milk in TBST (Tris-buffered saline-0.1 % Tween) at 25 ◦C for 3 h and then incubated with the following primary antibodies: rabbit polyclonal anti-GAPDH antibody (1:1000, Abcam, ab8245), rabbit monoclonal anti-CD63 antibody (1:1500, Abcam, ab217345), mouse monoclonal anti-CD81 antibody (1:1200, Abcam, ab79559), rabbit monoclonal anti-TSG101 antibody (1:2500, Abcam, ab125011), rabbit monoclonal anti-Alix antibody (1:2000, Abcam, ab186492), Rabbit polyclonal anti-MAT2A antibody (1:2500, Abcam, ab154343), Rabbit monoclonal anti-COX-2 antibody (1:2000, Abcam, ab15191), Rabbit monoclonal anti-MCP-1 antibody (1:1000, Abcam, ab214819), Rabbit monoclonal anti-iNOS antibody (1:800, Abcam, ab178945), rabbit monoclonal anti-STAT3 antibody (1:1500, Abcam, ab68153), rabbit monoclonal anti-AKT (phospho Y705) antibody (1:5000, Abcam, ab76315).

Techniques: Expressing, Incubation, Concentration Assay, Over Expression, Plasmid Preparation, Transfection, Western Blot, MTT Assay, Flow Cytometry, Control

Journal: Brain research bulletin

Article Title: MiR-26b-5p-modified hUB-MSCs derived exosomes attenuate early brain injury during subarachnoid hemorrhage via MAT2A-mediated the p38 MAPK/STAT3 signaling pathway.

doi: 10.1016/j.brainresbull.2021.07.014

Figure Lengend Snippet: Fig. 6. MiR-26b-5p-modified exosomes inhibited p38 MAPK/STAT3 signaling pathway to alleviate oxyHb-induced cell injury. PC12 cells were stimulated with 50 ng/mL oxyHb (cell density was 1 × 106) for 6 h and incubated with exosomes at a concentration of 100 μg/mL. Then, cells were transfected with miR-26b-5p inhibitor alone or together with si-MAT2A, respectively. A. Western botting was used to detect the protein expression of MAT2A as well as the phosphorylation levels of p38 MAPK and STAT3 proteins. B. MTT assay was used to detected cell proliferation. C. The protein expression of COX-2, MCP-1 and iNOS was analyzed by Western botting. GAPDH was used as an internal reference. N = 6, *P < 0.01.

Article Snippet: Then, equal amounts of proteins (15 μg/lane) were separated on a 12 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenedifluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA).The membranes were blocked in 5% (w/v) nonfat dry milk in TBST (Tris-buffered saline-0.1 % Tween) at 25 ◦C for 3 h and then incubated with the following primary antibodies: rabbit polyclonal anti-GAPDH antibody (1:1000, Abcam, ab8245), rabbit monoclonal anti-CD63 antibody (1:1500, Abcam, ab217345), mouse monoclonal anti-CD81 antibody (1:1200, Abcam, ab79559), rabbit monoclonal anti-TSG101 antibody (1:2500, Abcam, ab125011), rabbit monoclonal anti-Alix antibody (1:2000, Abcam, ab186492), Rabbit polyclonal anti-MAT2A antibody (1:2500, Abcam, ab154343), Rabbit monoclonal anti-COX-2 antibody (1:2000, Abcam, ab15191), Rabbit monoclonal anti-MCP-1 antibody (1:1000, Abcam, ab214819), Rabbit monoclonal anti-iNOS antibody (1:800, Abcam, ab178945), rabbit monoclonal anti-STAT3 antibody (1:1500, Abcam, ab68153), rabbit monoclonal anti-AKT (phospho Y705) antibody (1:5000, Abcam, ab76315).

Techniques: Modification, Incubation, Concentration Assay, Transfection, Western Blot, Expressing, Phospho-proteomics, MTT Assay

Journal: Brain research bulletin

Article Title: MiR-26b-5p-modified hUB-MSCs derived exosomes attenuate early brain injury during subarachnoid hemorrhage via MAT2A-mediated the p38 MAPK/STAT3 signaling pathway.

doi: 10.1016/j.brainresbull.2021.07.014

Figure Lengend Snippet: Fig. 7. MiR-26b-5p-modified exosomes attenuate early brain injury in SAH rats. The animal model of SAH was established by the classical occipital cistern secondary injection method. Thirty SD rats were divided into five groups on average. The cerebellar medullary cistern of rats in sham operation group was injected with normal saline. A. Relative expression of miR-26b-5p was detected by RT-qPCR. B. Western botting was used to detect the protein expression of MAT2A as well as the phosphorylation levels of p38 MAPK and STAT3 proteins. C. Behavioral and neurological functions of SAH rats were evaluated 24 h after modeling. D. Brain edema content in SAH rats was detected 24 h after modeling. E. The protein expression of COX-2, MCP-1 and iNOS was detected by Western botting. GAPDH was used as an invariant internal control for calculating protein-fold changes. N = 6, *P < 0.01.

Article Snippet: Then, equal amounts of proteins (15 μg/lane) were separated on a 12 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidenedifluoride (PVDF) membranes (Bio-Rad, Hercules, CA, USA).The membranes were blocked in 5% (w/v) nonfat dry milk in TBST (Tris-buffered saline-0.1 % Tween) at 25 ◦C for 3 h and then incubated with the following primary antibodies: rabbit polyclonal anti-GAPDH antibody (1:1000, Abcam, ab8245), rabbit monoclonal anti-CD63 antibody (1:1500, Abcam, ab217345), mouse monoclonal anti-CD81 antibody (1:1200, Abcam, ab79559), rabbit monoclonal anti-TSG101 antibody (1:2500, Abcam, ab125011), rabbit monoclonal anti-Alix antibody (1:2000, Abcam, ab186492), Rabbit polyclonal anti-MAT2A antibody (1:2500, Abcam, ab154343), Rabbit monoclonal anti-COX-2 antibody (1:2000, Abcam, ab15191), Rabbit monoclonal anti-MCP-1 antibody (1:1000, Abcam, ab214819), Rabbit monoclonal anti-iNOS antibody (1:800, Abcam, ab178945), rabbit monoclonal anti-STAT3 antibody (1:1500, Abcam, ab68153), rabbit monoclonal anti-AKT (phospho Y705) antibody (1:5000, Abcam, ab76315).

Techniques: Modification, Animal Model, Injection, Saline, Expressing, Quantitative RT-PCR, Western Blot, Phospho-proteomics, Control

Journal: eLife

Article Title: SAM homeostasis is regulated by CFI m -mediated splicing of MAT2A

doi: 10.7554/eLife.64930

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal anti-MAT2A , Novus , Cat#: NB110-94158; RRID: AB_1237164 , (1:2000).

Techniques: Recombinant, Plasmid Preparation, Mutagenesis, Binding Assay, Clone Assay, CRISPR, Knock-Out, Sequencing, Northern Blot, Negative Control, Software

Journal: Cell metabolism

Article Title: Transsulfuration activity can support cell growth upon extracellular cysteine limitation

doi: 10.1016/j.cmet.2019.09.009

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-MAT2A , Bethyl Laboratories , Cat# A304-279A-M; RRID: AB_2620475.

Techniques: Virus, Recombinant, SYBR Green Assay, Plasmid Preparation, Software, CRISPR