Journal: International Journal of Molecular Sciences

Article Title: Preparation of CPD Photolyase Nanoliposomes Derived from Antarctic Microalgae and Their Effect on UVB-Induced Skin Damage in Mice

doi: 10.3390/ijms232315148

Figure Lengend Snippet: The effect of CPDNL on mRNA expression of MAPK/AP-1/MMP signaling pathways in UVB-induced skin. ( a–c ) The mRNA expression of ERK, JNK, and P38; ( d ) the mRNA expression of AP-1; ( e , f ) the mRNA expression of MMP-1 and MMP-2. Data are presented as mean ± SEM. ** p < 0.01 and *** p < 0.001.

Article Snippet: The membrane was incubated with the primary antibody against NF-κВ (BA1297-2, Boster, Wuhan, China), TNF-α (BA14901, Boster), IL-6 (BA4339-2, Boster), COX-2 (BA3708, Boster), ERK (GB112238, Servicebio, Wuhan, China), JNK (M02608-3, Boster), P38 (A00176-2, Boster), AP-1 (GB11270, Servicebio), MMP-1 (A00733-1, Boster), and MMP-2 (A00286, Boster) at 4 °C overnight, followed by washing at TBST, and then incubated with goat anti-rabbit HRP (horseradish peroxidase)-IgG (BA1055, Boster) as the secondary antibody for 45 min. After washed with TBST 3 times for 10 min, membrane was incubated in SuperSignal ECL Western substrate (Biosharp, Hefei, China).

Techniques: Expressing, Protein-Protein interactions

Journal: International Journal of Molecular Sciences

Article Title: Preparation of CPD Photolyase Nanoliposomes Derived from Antarctic Microalgae and Their Effect on UVB-Induced Skin Damage in Mice

doi: 10.3390/ijms232315148

Figure Lengend Snippet: The effect of CPDNL on protein level of MAPK/AP-1/MMP signaling pathways in UVB-induced skin. The protein expression level ( a ) of ERK ( b ), JNK ( c ), P38 ( d ), AP-1 ( e ), MMP-1 ( f ), and MMP-2 ( g ) was determined by Western blot analysis. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The membrane was incubated with the primary antibody against NF-κВ (BA1297-2, Boster, Wuhan, China), TNF-α (BA14901, Boster), IL-6 (BA4339-2, Boster), COX-2 (BA3708, Boster), ERK (GB112238, Servicebio, Wuhan, China), JNK (M02608-3, Boster), P38 (A00176-2, Boster), AP-1 (GB11270, Servicebio), MMP-1 (A00733-1, Boster), and MMP-2 (A00286, Boster) at 4 °C overnight, followed by washing at TBST, and then incubated with goat anti-rabbit HRP (horseradish peroxidase)-IgG (BA1055, Boster) as the secondary antibody for 45 min. After washed with TBST 3 times for 10 min, membrane was incubated in SuperSignal ECL Western substrate (Biosharp, Hefei, China).

Techniques: Protein-Protein interactions, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Preparation of CPD Photolyase Nanoliposomes Derived from Antarctic Microalgae and Their Effect on UVB-Induced Skin Damage in Mice

doi: 10.3390/ijms232315148

Figure Lengend Snippet: Real-time quantitative PCR primer sequences.

Article Snippet: The membrane was incubated with the primary antibody against NF-κВ (BA1297-2, Boster, Wuhan, China), TNF-α (BA14901, Boster), IL-6 (BA4339-2, Boster), COX-2 (BA3708, Boster), ERK (GB112238, Servicebio, Wuhan, China), JNK (M02608-3, Boster), P38 (A00176-2, Boster), AP-1 (GB11270, Servicebio), MMP-1 (A00733-1, Boster), and MMP-2 (A00286, Boster) at 4 °C overnight, followed by washing at TBST, and then incubated with goat anti-rabbit HRP (horseradish peroxidase)-IgG (BA1055, Boster) as the secondary antibody for 45 min. After washed with TBST 3 times for 10 min, membrane was incubated in SuperSignal ECL Western substrate (Biosharp, Hefei, China).

Techniques: Real-time Polymerase Chain Reaction

Journal: Frontiers in Pharmacology

Article Title: Induction of Mkp-1 and Nuclear Translocation of Nrf2 by Limonoids from Khaya grandifoliola C.DC Protect L-02 Hepatocytes against Acetaminophen-Induced Hepatotoxicity

doi: 10.3389/fphar.2017.00653

Figure Lengend Snippet: Active sub-fractions and isolated limonoids prevented APAP-induced JNK activation. Cells were treated without or with APAP (10 mM), or co-treated with APAP (10 mM) and active sub-fractions or silymarin (100 μg/mL), isolated limonoids (40 μM) or JNK inhibitor (20 μM) for 6 h. After treatment, total proteins were extracted from cells and the lysates were probed for JNK2 and p-JNK by western blotting. β-actin was used as loading control. Each blot represents one of three independent experiments. (A,C) Effect of active sub-fractions and isolated compounds on JNK activation, respectively. (B,D) Densitometry analysis of blots, respectively, for active sub-fractions and isolated compounds. Values are means ± SD of three independent experiments in triplicate. ANOVA analysis: F (6,14) = 322.0, P < 0.0001 (B) and F (5,12) = 1162.0, P < 0.0001 (D) . Δ Values significantly different compared to control group ( P < 0.05); ∗ values significantly different compared to APAP intoxicated group ( P < 0.05) using Bonferroni’s test. Lane1: control; Lane2: APAP; Lane3: silymarin+APAP; Lane4: KgF25+APAP; Lane5: KgF25sf1+APAP; Lane6: KgF25sf2+APAP; Lane7: Kgf25sf3+APAP. Sil: silymarin; KgF25: methylene chloride/methanol (75:25, v/v) fraction of K. grandifoliola ; KgF25sf1: sub-fraction 1 of KgF25; KgF25sf2: sub-fraction 2 of KgF25; KgF25sf3: sub-fraction 3 of KgF25; SP: JNK inhibitor SP600125; C-A: 17-epi-methyl-6-hydroxyangolensate; C-B: 7-deacetoxy-7-oxogedunin; C-C: 7-deacetoxy-7R-hydroxygedunin.

Article Snippet: Rabbit polyclonal anti-CYP2E1 antibody (1:1500 dilution) was purchased from Sino Biological Inc. (Beijing, China); Rabbit polychonal anti-phospho-JNK1/JNK2 and JNK2 antibodies (1:1000 dilution) were purchased from Signalway Antibody (Baltimore, MD, United States); Rabbit polyclonal anti-MKP-1, Nrf2, Keap-1, AIF, COX IV, Lamin B and mouse polyclonal anti-Bax antibodies (all 1:1000 dilution) were purchased from Beijing Biosynthesis Biotechnology CO., LTD. (Beijing, China); Mouse monoclonal anti-βactin primary antibody (1:5000 dilution), horseradish peroxidase-conjugated goat anti-rabbit and anti-mouse IgG AP-linked secondary antibodies (1:2000 dilution) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States); TRIzol ® Reagent was purchased from Ambion Life Technologies (Carlsbad, CA, United States); First-Strand cDNA Synthesis Kit was purchased from Promega (Madison, WI, United States); iTaq Universal SYBR Green Supermix Kit was purchased from Bio-Rad Laboratories (Hercules, CA, United States); All primers of the genes of interest were synthetized by TSINGKE Biological Technology Company (Beijing, China).

Techniques: Isolation, Activation Assay, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Induction of Mkp-1 and Nuclear Translocation of Nrf2 by Limonoids from Khaya grandifoliola C.DC Protect L-02 Hepatocytes against Acetaminophen-Induced Hepatotoxicity

doi: 10.3389/fphar.2017.00653

Figure Lengend Snippet: Limonoids from K. grandifoliola prevented mitochondrial p-JNK translocation. Cells were treated without or with APAP (10 mM), or co-treated with APAP (10 mM) and isolated limonoids (40 μM) or JNK inhibitor (20 μM) for 6 and 12 h. After treatment, p-JNK level was detected into the cytosolic and mitochondrial fractions by western blotting. JNK2 was used as internal control. Each blot represents one of three independent experiments. (A,B) Effect of isolated compounds on JNK activation, respectively, at 6 and 12 h after treatment. (C) densitometry analysis of blots. Values are means ± SD of three independent experiments in triplicate. ANOVA analysis: F (5,12) = 1119.0, P < 0.0001 and F (5,12) = 1841.0, P < 0.0001 (C) . Δ Values significantly different compared to control group ( P < 0.05); ∗ values significantly different compared to APAP intoxicated group ( P < 0.05) using Bonferroni’s test. SP: JNK inhibitor SP600125; C-A: 17-epi -methyl-6-hydroxyangolensate; C-B: 7-deacetoxy-7-oxogedunin; C-C: 7-deacetoxy-7R-hydroxygedunin.

Article Snippet: Rabbit polyclonal anti-CYP2E1 antibody (1:1500 dilution) was purchased from Sino Biological Inc. (Beijing, China); Rabbit polychonal anti-phospho-JNK1/JNK2 and JNK2 antibodies (1:1000 dilution) were purchased from Signalway Antibody (Baltimore, MD, United States); Rabbit polyclonal anti-MKP-1, Nrf2, Keap-1, AIF, COX IV, Lamin B and mouse polyclonal anti-Bax antibodies (all 1:1000 dilution) were purchased from Beijing Biosynthesis Biotechnology CO., LTD. (Beijing, China); Mouse monoclonal anti-βactin primary antibody (1:5000 dilution), horseradish peroxidase-conjugated goat anti-rabbit and anti-mouse IgG AP-linked secondary antibodies (1:2000 dilution) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States); TRIzol ® Reagent was purchased from Ambion Life Technologies (Carlsbad, CA, United States); First-Strand cDNA Synthesis Kit was purchased from Promega (Madison, WI, United States); iTaq Universal SYBR Green Supermix Kit was purchased from Bio-Rad Laboratories (Hercules, CA, United States); All primers of the genes of interest were synthetized by TSINGKE Biological Technology Company (Beijing, China).

Techniques: Translocation Assay, Isolation, Western Blot, Activation Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: A novel function and mechanism of ischemia-induced retinal astrocyte-derived exosomes for RGC apoptosis of ischemic retinopathy

doi: 10.1016/j.omtn.2024.102209

Figure Lengend Snippet: miR-329-5p protected the retina from ischemic-reperfusion injury by targeting the MAPK pathway (A and B) Venn diagrams showing the intersection of predicted target genes of hsa-miR-329-5p and mmu-miR-329-5p, and MAPK8 (Jnk1) is a potential gene targeted by both miRNAs. (C) Real-time qPCR analysis of MAPK8 mRNA expression after intravitreal treatment of PBS, scrambled miRNA (miR-NC), and miR-329-5p (n = 5). (D and E) WB and quantification of protein expressions of MAPK8- related pathway on retinal tissues after intravitreal treatments of PBS, miR-NC, and miR-329-5p (n = 5). (F) IHC of MAPK8 expression on retinal tissues after various intravitreal treatments (n = 5 animals; scale bar, 100 μm). (G) Dual-luciferase reporter gene plasmid design and the predicted target sites. (H) The dual luciferase reporter gene assay showed that miR-329-5p significantly inhibited the luciferase activity of the MAPK8 -WT1 3′UTR reporter gene in 293T cells. Data was presented as mean ± SD, n = 3. Student’s t test was used for statistical analysis. Data were statistically analyzed by one-way ANOVA followed by Tukey’s post hoc analysis. Statistical significance at ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. ns, not statistically significant.

Article Snippet: Anti- p -JNK1 (Thr183/Tyr185) rabbit pAb , Abcepta, Cat#. AP22143a.

Techniques: Expressing, Luciferase, Plasmid Preparation, Reporter Gene Assay, Activity Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: A novel function and mechanism of ischemia-induced retinal astrocyte-derived exosomes for RGC apoptosis of ischemic retinopathy

doi: 10.1016/j.omtn.2024.102209

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: Anti- p -JNK1 (Thr183/Tyr185) rabbit pAb , Abcepta, Cat#. AP22143a.

Techniques:

Journal: Molecular Therapy. Nucleic Acids

Article Title: A novel function and mechanism of ischemia-induced retinal astrocyte-derived exosomes for RGC apoptosis of ischemic retinopathy

doi: 10.1016/j.omtn.2024.102209

Figure Lengend Snippet: Primer sequences used in real-time qPCR test

Article Snippet: Anti- p -JNK1 (Thr183/Tyr185) rabbit pAb , Abcepta, Cat#. AP22143a.

Techniques: Sequencing