Journal: American Journal of Translational Research

Article Title: lncRNA PART1 mitigates MPP + -induced neuronal injury in SH-SY5Y cells via micRNA-106b-5p/MCL1 axis

doi:

Figure Lengend Snippet: The sequence of reagents used in cell transfection

Article Snippet: To examine whether microRNA-106b-5p targeted PART1 or MCL1 directly, PART1-WT, PART1-MUT, MCL1-WT and MCL1-MUT reporter vectors were purchased from Promega (Madison, WI, USA).

Techniques: Sequencing

Journal: American Journal of Translational Research

Article Title: lncRNA PART1 mitigates MPP + -induced neuronal injury in SH-SY5Y cells via micRNA-106b-5p/MCL1 axis

doi:

Figure Lengend Snippet: The sequence of primers

Article Snippet: To examine whether microRNA-106b-5p targeted PART1 or MCL1 directly, PART1-WT, PART1-MUT, MCL1-WT and MCL1-MUT reporter vectors were purchased from Promega (Madison, WI, USA).

Techniques: Sequencing

Journal: American Journal of Translational Research

Article Title: lncRNA PART1 mitigates MPP + -induced neuronal injury in SH-SY5Y cells via micRNA-106b-5p/MCL1 axis

doi:

Figure Lengend Snippet: The expression of PART1 in PD. A: The protein expressions of TH and α-synuclein were examined by western blot in the mouse model of PD. β-actin was used as lading control; B: The expression of PART1 in brain tissues was examined in the mouse model of PD by RT-qPCR; C: The expression of PART1 in SH-SY5Y cells treated with MPP+ was examined by RT-qPCR. **P<0.01. PD: Parkinson’s disease; MPP+: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine.

Article Snippet: To examine whether microRNA-106b-5p targeted PART1 or MCL1 directly, PART1-WT, PART1-MUT, MCL1-WT and MCL1-MUT reporter vectors were purchased from Promega (Madison, WI, USA).

Techniques: Expressing, Western Blot, Quantitative RT-PCR

Journal: American Journal of Translational Research

Article Title: lncRNA PART1 mitigates MPP + -induced neuronal injury in SH-SY5Y cells via micRNA-106b-5p/MCL1 axis

doi:

Figure Lengend Snippet: Overexpression of PART1 alleviated neuronal injury. SH-SY5Y cells transfected with pcDNA3.1-NC or PART1 pcDNA3.1 were treated with 1 μM MPP+ for 48 h. A: The expression of PART1 was examined by RT-qPCR; B: The cell viability was assessed by CCK8 assay; C: The cell apoptosis rate was detected by Annexin V-FITC/PI assay; D: The expression of cleaved caspase 3 was detected by western blot; E: The activity of cleaved caspase 3 was detected; F-H: The levels of inflammatory cytokines were examined by ELISA; I-K: The levels of LDH, ROS and SOD were examined. *P<0.05, **P<0.01. MPP+: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; LDH: Lactate dehydrogenase; ROS: Reactive oxygen; SOD: Superoxide dismutase.

Article Snippet: To examine whether microRNA-106b-5p targeted PART1 or MCL1 directly, PART1-WT, PART1-MUT, MCL1-WT and MCL1-MUT reporter vectors were purchased from Promega (Madison, WI, USA).

Techniques: Over Expression, Transfection, Expressing, Quantitative RT-PCR, CCK-8 Assay, Western Blot, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Translational Research

Article Title: lncRNA PART1 mitigates MPP + -induced neuronal injury in SH-SY5Y cells via micRNA-106b-5p/MCL1 axis

doi:

Figure Lengend Snippet: PART1 targets microRNA-106b-5p directly. (A) The predictive binding site of PART1 was predicted by Starbase 2.0; (B) The expression of microRNA-106b-5p was examined by RT-qPCR in SH-SY5Y cells transfected with PART1 pcDNA3.1; (C) The expression of microRNA-106b-5p was examined by RT-qPCR in the SH-SY5Y cells transfected with mimics NC and microRNA-106b-5p mimics; (D) The DLR assay was used to detect the binding relationship between microRNA-106b-5p and PART1 in SH-SY5Y cells co-transfected with PART1-WT or PART1-MUT and mimics NC or microRNA-106b-5p mimics; (E, F) The RIP assay (E) and RNA pull down assay (F) were used to examine the interaction between PART1 and microRNA-106b-5p in SH-SY5Y cells; (G, H) The expressions of microRNA-106b-5p in SH-SY5Y cells (G) and PD mouse model (H) were examined by RT-qPCR; (I) The correlation analysis of PART1 and microRNA-106b-5p in PD mouse model. **P<0.01. DLR: Dual-luciferase report; PD: Parkinson’s disease.

Article Snippet: To examine whether microRNA-106b-5p targeted PART1 or MCL1 directly, PART1-WT, PART1-MUT, MCL1-WT and MCL1-MUT reporter vectors were purchased from Promega (Madison, WI, USA).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Transfection, Pull Down Assay, Luciferase

Journal: American Journal of Translational Research

Article Title: lncRNA PART1 mitigates MPP + -induced neuronal injury in SH-SY5Y cells via micRNA-106b-5p/MCL1 axis

doi:

Figure Lengend Snippet: PART1 attenuated the injury of SH-SY5Y cells induced by MPP+ via microRNA-106b-5p/MCL1 axis. After transfection with PART1 pcDNA3.1, microRNA-106b-5p mimics, and MCL1 siRNA, SH-SY5Y cells were treated with 1 μM MPP+ for 48 h. A: The expression of MCL1 was examined by western blot; B: The cell viability was assessed by CCK8 assay; C: The cell apoptosis rate was detected by Annexin V-FITC/PI assay; D: The expression of cleaved caspase 3 was detected by western blot; E: The activity of cleaved caspase 3 was detected; F-H: The levels of inflammatory cytokines were examined by ELISA; I-K: The levels of LDH, ROS and SOD were examined. *P<0.05, **P<0.01. MPP+: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine.

Article Snippet: To examine whether microRNA-106b-5p targeted PART1 or MCL1 directly, PART1-WT, PART1-MUT, MCL1-WT and MCL1-MUT reporter vectors were purchased from Promega (Madison, WI, USA).

Techniques: Transfection, Expressing, Western Blot, CCK-8 Assay, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Translational Research

Article Title: lncRNA PART1 mitigates MPP + -induced neuronal injury in SH-SY5Y cells via micRNA-106b-5p/MCL1 axis

doi:

Figure Lengend Snippet: The schematic diagram of PART1/microRNA-106b-5p/MCL1 axis.

Article Snippet: To examine whether microRNA-106b-5p targeted PART1 or MCL1 directly, PART1-WT, PART1-MUT, MCL1-WT and MCL1-MUT reporter vectors were purchased from Promega (Madison, WI, USA).

Techniques:

Journal: Journal of visualized experiments : JoVE

Article Title: Establishing in situ closed circuit perfusion of lower abdominal organs and hind limbs in mice

doi: 10.3791/60847

Figure Lengend Snippet: Equipment and Materials

Article Snippet: ​ table ft1 table-wrap mode="anchored" t5 caption a7 Name of Material/ Equipment Company Catalog Number Comments/Description Equipment SCP plugsys servo control F/Perfusion (part 1 in ) Harvard Apparatus 732806 control the purfusion speed Thermocirculator (part 2 in ) Harvard Apparatus 724927 circulating water bath for all water-jacketed components Moist chamber cover with quartz window (part 3 in ) Harvard Apparatus 733524 keep the chamber’s temperature Roller pump (part 4 in ) Harvard Apparatus 730113 deliver perfusate to cannula in the moist chamber Three-way stopcock (part 5 in ) Cole-Palmer 30600-02 Moist chamber bubble trap (part 6 in ) Harvard Apparatus 733692 Customized as the perfisate container; also enabled constant pressure perfusion Silicone tubing set (arrows in ) Harvard Apparatus (TYGON) 733456 Moist chamber with metal tube heat exchanger Harvard Apparatus 732901 Water-jacketed moist chamber with lid to maintain perfusate and mouse temperature Silicone pad Harvard Apparatus Table for moist chamber Harvard Apparatus 734198 Veterinary anesthesia machine Highland HME109 Carbon dioxide, USP Airgas healthcare 19087-5283 Oxygen compressed, USP Airgas healthcare C2649150AE06 3.5X-90X stereo zoom microscope on boom stand with LED light Amscope SKU: SM-3BZ-80S Confocal microscope NIKON ECLIPSE Ti2 Disposable Sterile Cautery Pen with High Temp FIAB F7244 Olsen-Hegar needle holders with suture cutters Fine Science Tools (FST) 125014 Student standard pattern forceps Fine Science Tools (FST) 91100-12 Surgical Scissors Fine Science Tools (FST) 14001-14 Materials Permanent marker Staedtler 342-9 Filter (0.2 μm) ThermoFisher 42225-CA Filter for 5% BSA-RINGER’S Syringe (60ml) BD 309653 Filter for 5% BSA-RINGER’S Syringe (10 ml) Fisher Scientific 14-823-2E 19-G BD PrecisionGlide needle BD 305186 For immobilizing the Insyte Autoguard Winged needle and scratching the cortical bone 4-0 silk sutures Keebomed-Hopemedical 427411 6-0 silk sutures Keebomed-Hopemedical 427401 Reagents Isoflurane Piramal Enterprises Limited 66794-017-25 Ethanol (70% ( vol/vol )) Pharmco 111000190 Baxter Viaflex lactate Ringer’s EMRN Medical Supplies Inc. JB2324 Bovine serum albumin Thermo Fisher 11021-037 Hoechst33342 Life Technologies H3570 DyLight-649-lectin Vector Laboratories,Inc.

Techniques: Microscopy, Marker, Plasmid Preparation, Mouse Assay