Journal: Oxidative Medicine and Cellular Longevity

Article Title: Methane Ameliorates Lipopolysaccharide-Induced Acute Orchitis by Anti-inflammatory, Antioxidative, and Antiapoptotic Effects via Regulation of the PK2/PKR1 Pathway

doi: 10.1155/2020/7075836

Figure Lengend Snippet: Sequences of primers utilized for RT-qPCR analysis.

Article Snippet: Membranes were blocked in 5% skimmed milk/PBS for 1 h at 4°C and incubated with the following primary antibodies: rabbit polyclonal anti-PK2 (cat. no. ab76747, 1 : 200, Abcam), rabbit polyclonal anti-PKR1 (cat. no. APR-041, 1 : 200, Alomone Labs), and mouse monoclonal anti-GAPDH (cat no. WB2197, 1 : 2000, Well Biotech Co., Ltd.) at 4°C overnight and then incubated with horseradish peroxidase- (HRP-) labeled goat anti-rabbit secondary antibody (cat. no. 111-035-003, 1 : 5000, Jackson ImmunoResearch Laboratories, Inc.) at 4°C for 1 h.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Methane Ameliorates Lipopolysaccharide-Induced Acute Orchitis by Anti-inflammatory, Antioxidative, and Antiapoptotic Effects via Regulation of the PK2/PKR1 Pathway

doi: 10.1155/2020/7075836

Figure Lengend Snippet: Methane decreased the expression of PKR1 in testes of rats with LPS-induced acute orchitis. RT-qPCR (a), WB (b-c), and IHC (d-g) were used to determine the expression of PKR1 in the testes at 72 h after LPS injection. ∗ P < 0.05 vs the Control group, # P < 0.05 vs the MS group, $ P < 0.05 vs the LPS group. Scale bar = 50 μ m.

Article Snippet: Membranes were blocked in 5% skimmed milk/PBS for 1 h at 4°C and incubated with the following primary antibodies: rabbit polyclonal anti-PK2 (cat. no. ab76747, 1 : 200, Abcam), rabbit polyclonal anti-PKR1 (cat. no. APR-041, 1 : 200, Alomone Labs), and mouse monoclonal anti-GAPDH (cat no. WB2197, 1 : 2000, Well Biotech Co., Ltd.) at 4°C overnight and then incubated with horseradish peroxidase- (HRP-) labeled goat anti-rabbit secondary antibody (cat. no. 111-035-003, 1 : 5000, Jackson ImmunoResearch Laboratories, Inc.) at 4°C for 1 h.

Techniques: Expressing, Quantitative RT-PCR, Injection

Journal: Scientific Reports

Article Title: Bv8/prokineticin 2 is involved in Aβ-induced neurotoxicity

doi: 10.1038/srep15301

Figure Lengend Snippet: Relative PROK2, PKR 1 and PKR 2 mRNA levels were determined by qRT-PCR in CNs (1 × 10 6 cells/well) treated at 12 DIV with Aβ 1–42 (20 μM) for different times ( A – C ), in cortex ( D – F ) and hippocampus ( G – I ) from Aβ 1–42 injected rats at different time points. The mRNA expression levels were expressed in relation to β-actina and presented as fold of increase relative to control cultures or Sham rats. Data represent means (±SEM) from at least 4 independent experimental points run in triplicate and statistically significant differences were calculated by one-way analysis of variance (ANOVA) for repeated measures followed by Bonferroni’s test for multiple comparisons (*p < 0.05, **p < 0.01 vs CTR or sham).

Article Snippet: Living mixed cortical cells were stained for 1 h at 4 °C using rabbit polyclonal anti-PKR 1 or anti-PKR 2 antibodies 1:400 in PBS (Alomone Labs, Jerusalem, Israel), washed in PBS and incubated with a goat anti-rabbit rhodamine-conjugated secondary antibody (Sigma) for 30 min at room temperature.

Techniques: Quantitative RT-PCR, Injection, Expressing

Journal: Scientific Reports

Article Title: Bv8/prokineticin 2 is involved in Aβ-induced neurotoxicity

doi: 10.1038/srep15301

Figure Lengend Snippet: Representative confocal images of cultured mixed cortical neurons (CNs) stained with anti-PROK2 ( A – D ) or anti PKR 1 ( E – H ) or anti PKR2 antibodies ( I – L ) (red) in control conditions (CTRL) and after 48 h Aβ 1–42 treatment (Aβ). Neurons were stained with NeuN (green), astrocytes with GFAP (green), and nuclei with Hoecsht (blue). Panel ( A–D ) PROK2 low to medium immunoreactivity was found in neuronal cell bodies and astrocytes processes (arrows) in control conditions (first row) which increased after Aβ 1–42 treatment mainly in the cell bodies (second row: arrow). Panel ( E–H ) PKR 1 immunoreactivity was undetectable (first row) under control conditions in both neurons and astrocytes. After Aβ 1–42 treatment PKR 1 immunoreactivity displayed a selective neuronal increase confined to putative endoplasmic reticulus-Golgi complex (arrows) while astrocytes were still devoid of staining (arrowheads). Please note, close to astrocytes there are some PKR 1 positive neurons (arrows). Panel ( I–L ) Very low PKR 2 immunoreactivity was detected under control conditions while, after Aβ 1–42 treatment increased in neuronal cell bodies and astrocyte processes (arrows). Note the selective increase in a dying cell body surrounded by astrocytic processes (arrowheads). Scale bar: 15 μm.

Article Snippet: Living mixed cortical cells were stained for 1 h at 4 °C using rabbit polyclonal anti-PKR 1 or anti-PKR 2 antibodies 1:400 in PBS (Alomone Labs, Jerusalem, Israel), washed in PBS and incubated with a goat anti-rabbit rhodamine-conjugated secondary antibody (Sigma) for 30 min at room temperature.

Techniques: Cell Culture, Staining