Journal: American journal of physiology. Heart and circulatory physiology

Article Title: Loss of cardiac sympathetic neurotransmitters in heart failure and NE infusion is associated with reduced NGF.

doi: 10.1152/ajpheart.00319.2001

Figure Lengend Snippet: Fig. 2. Representative Western blots showing reductions of left ven- tricular tyrosine kinase A (TrKA) protein in CHF (A) and NE-treated dogs (B) compared with sham-operated control and saline-treated animals. PC12 cell lysates were used as a positive control for TrKA shown in B.

Article Snippet: To determine the specificity of the primary antibodies for NGF and TrKA, we used PC12 cell lysates (Santa Cruz Biotechnology) as a positive control and nonimmune rabbit serum (Oncogene Research Products; Cambridge, MA) as negative control.

Techniques: Western Blot, Control, Saline, Positive Control

Journal: Bioconjugate chemistry

Article Title: Simple Method To Prepare Oligonucleotide-Conjugated Antibodies and Its Application in Multiplex Protein Detection in Single Cells.

doi: 10.1021/acs.bioconjchem.5b00613

Figure Lengend Snippet: Fig. 6. Detection of proteins in cell lysates. A panel of 24-plex antibody binders was used to detect their respective target proteins in MCF7, K562, A549, and A431 cell lysates. The cell lysates were diluted to different concentrations and reacted with a mixture of antibody binders. Only selected examples of protein targets CSTB (A), CASP3 (B), Ki-67(C), and GATA3 (D) are shown. Ct values were shown as the means ± SD. Ct: cycle threshold.

Article Snippet: MCF7, K562, A549, and A431 cell lysates were obtained from Novus (Littleton, CO).

Techniques:

Journal: Cells

Article Title: Melatonin Attenuates Spinal Cord Injury in Mice by Activating the Nrf2/ARE Signaling Pathway to Inhibit the NLRP3 Inflammasome

doi: 10.3390/cells11182809

Figure Lengend Snippet: The oxidative damage scavenging effect of melatonin in neuron cells. ( A ) The survival rates of PC12 cells that received different concentrations of melatonin treatment were detected using a CCK-8 kit (n = 8). ( B ) The ROS produced in PC12 cells that received different treatments were detected by ROS staining (n = 6, scale bar = 100 µm). ( C , D ) The mitochondrial membrane potential in PC12 cells treated with drugs was detected by JC-1 staining (n = 8, scale bar = 100 µm). ( E – G ) The contents of SOD and MDA and the activity of GSH-PX were detected using the corresponding kits in PC12 cells treated with different treatment factors (n = 10). Data are shown as the means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of IL-1β and IL-18 were measured in the PC12 cell culture medium using ELISA kits according to the manufacturer’s instructions (Elabscience, China).

Techniques: CCK-8 Assay, Produced, Staining, Membrane, Activity Assay

Journal: Cells

Article Title: Melatonin Attenuates Spinal Cord Injury in Mice by Activating the Nrf2/ARE Signaling Pathway to Inhibit the NLRP3 Inflammasome

doi: 10.3390/cells11182809

Figure Lengend Snippet: Melatonin suppresses the NLRP3 inflammasome in PC12 cells treated with H2O2 by activating the Nrf2/ARE pathway. ( A – I ) Western blot detection and statistical analysis of NLRP3-inflammasome-related proteins NLRP3, caspase-1, ASC, and IL-1β; and antioxidant-stress-related proteins nuclear Nrf2, HO-1, and NQO-1 in PC12 cells from each group (n = 6). ( J , K ) The levels of IL-1β and IL-18 in the supernatant of PC12 cells were determined by ELISA kits (n = 6). Data are shown as the means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of IL-1β and IL-18 were measured in the PC12 cell culture medium using ELISA kits according to the manufacturer’s instructions (Elabscience, China).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Melatonin Attenuates Spinal Cord Injury in Mice by Activating the Nrf2/ARE Signaling Pathway to Inhibit the NLRP3 Inflammasome

doi: 10.3390/cells11182809

Figure Lengend Snippet: Immunofluorescence double staining of Nrf2 and NLRP3 in PC12 cells treated with H2O2. Immunofluorescence double staining was performed to test the protein level of Nrf2 ( A , B ) and NLRP3 ( C , D ) in PC12 cells from each group (n = 8, scale bar = 100 µm). Data are shown as the means ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of IL-1β and IL-18 were measured in the PC12 cell culture medium using ELISA kits according to the manufacturer’s instructions (Elabscience, China).

Techniques: Immunofluorescence, Double Staining

Journal: Molecular Medicine Reports

Article Title: Inflammatory response and oxidative stress attenuated by sulfiredoxin-1 in neuron-like cells depends on nuclear factor erythroid-2-related factor 2

doi: 10.3892/mmr.2020.11545

Figure Lengend Snippet: SRX1 expression levels are downregulated in PC12 cells stimulated with LPS. (A) Cell Counting Kit-8 assay was used to determine the cell viability of PC12 cells following the stimulation with a range of concentrations of LPS. (B) mRNA and (C) protein expression levels of SRX1 in PC12 cells stimulated with a range of concentrations of LPS were determined using reverse transcription-quantitative PCR and western blotting, respectively. Data are expressed as the mean ± SD from 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001. SRX1, sulfiredoxin-1; LPS, lipopolysaccharide.

Article Snippet: Rat pheochromocytoma PC12 cells (China Center for Type Culture Collection) were cultured in DMEM (Hyclone; Cytiva), supplemented with 10% FBS (Hyclone; Cytiva), and maintained at 37°C under humid conditions with 5% CO 2 and 95% air.

Techniques: Expressing, Cell Counting, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: Molecular Medicine Reports

Article Title: Inflammatory response and oxidative stress attenuated by sulfiredoxin-1 in neuron-like cells depends on nuclear factor erythroid-2-related factor 2

doi: 10.3892/mmr.2020.11545

Figure Lengend Snippet: Overexpression of SRX1 reduces the inflammatory response in PC12 cells. (A) Transfection efficiency of Ov-SRX1 in PC12 cells was determined using RT-qPCR. (B) mRNA and (C) protein expression levels of SRX1 in PC12 cells transfected with Ov-SRX1 and stimulated with 5 µg/ml LPS were determined using RT-qPCR and western blotting, respectively. (D) Cell Counting Kit-8 assay was used to determine the cell viability of PC12 cells transfected with Ov-SRX1 and stimulated with 5 µg/ml LPS. (E) TNF-α, IL-1β, IL-18 and IL-10 levels in PC12 cells transfected with Ov-SRX1 and stimulated with 5 µg/ml LPS were detected using ELISA kits. Data are expressed as the mean ± SD from 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001. SRX1, sulfiredoxin-1; LPS, lipopolysaccharide; Ov, overexpression; NC, negative control; RT-qPCR, reverse transcription-quantitative PCR; TNF-α, tumor necrosis factor α; IL, interleukin.

Article Snippet: Rat pheochromocytoma PC12 cells (China Center for Type Culture Collection) were cultured in DMEM (Hyclone; Cytiva), supplemented with 10% FBS (Hyclone; Cytiva), and maintained at 37°C under humid conditions with 5% CO 2 and 95% air.

Techniques: Over Expression, Transfection, Quantitative RT-PCR, Expressing, Western Blot, Cell Counting, Enzyme-linked Immunosorbent Assay, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Inflammatory response and oxidative stress attenuated by sulfiredoxin-1 in neuron-like cells depends on nuclear factor erythroid-2-related factor 2

doi: 10.3892/mmr.2020.11545

Figure Lengend Snippet: Overexpression of SRX1 relieves oxidative stress in PC12 cells. (A) Intracellular ROS in PC12 cells transfected with Ov-SRX1 and stimulated with 5 µg/ml LPS was stained using 2′,7′-dichlorofluorescin. Scale bar, 50 µm. (B) MDA content and (C) SOD activity in PC12 cells transfected with Ov-SRX1 and stimulated with 5 µg/ml LPS were analyzed using commercial detection kits. (D) Protein expression levels of antioxidative proteins PRDX1, PRDX6, TXNRD1 and SOD2 in PC12 cells transfected with Ov-SRX1 and stimulated with 5 µg/ml LPS were analyzed using western blotting. Data are expressed as the mean ± SD from 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001. SRX1, sulfiredoxin-1; LPS, lipopolysaccharide; Ov, overexpression; NC, negative control; ROS, reactive oxygen species; MDA, malondialdehyde; SOD, superoxide dismutase; PRDX, peroxiredoxin; TXNRD1, thioredoxin reductase 1.

Article Snippet: Rat pheochromocytoma PC12 cells (China Center for Type Culture Collection) were cultured in DMEM (Hyclone; Cytiva), supplemented with 10% FBS (Hyclone; Cytiva), and maintained at 37°C under humid conditions with 5% CO 2 and 95% air.

Techniques: Over Expression, Transfection, Staining, Activity Assay, Expressing, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: Inflammatory response and oxidative stress attenuated by sulfiredoxin-1 in neuron-like cells depends on nuclear factor erythroid-2-related factor 2

doi: 10.3892/mmr.2020.11545

Figure Lengend Snippet: NRF2 controls the expression levels of downstream target genes and SRX1. (A) Expression levels of nuclear NRF2 and downstream target genes NQO1 and HO-1 in PC12 cells stimulated with 5 µg/ml LPS were analyzed using western blotting. (B) mRNA expression levels of SRX1, NQO1 and HO-1 in PC12 cells stimulated with 5 µg/ml LPS with or without 25 µM TBHQ or 5 µM ML385 treatment were determined using reverse transcription-quantitative PCR. (C) Western blotting analysis was performed to analyze the expression levels of nuclear NRF2, NQO1, HO-1 and SRX1 in PC12 cells stimulated with 5 µg/ml LPS with or without 25 µM TBHQ or 5 µM ML385 treatment. Data are expressed as the mean ± SD from 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001. SRX1, sulfiredoxin-1; LPS, lipopolysaccharide; Ov, overexpression; NC, negative control; NRF2, nuclear factor erythroid-2-related factor 2; HO-1, heme oxygenase 1; NQO1, NAD(P)H dehydrogenase quinone 1; TBHQ, tert-butylhydroquinone.

Article Snippet: Rat pheochromocytoma PC12 cells (China Center for Type Culture Collection) were cultured in DMEM (Hyclone; Cytiva), supplemented with 10% FBS (Hyclone; Cytiva), and maintained at 37°C under humid conditions with 5% CO 2 and 95% air.

Techniques: Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Over Expression, Negative Control