Journal: Journal of Integrative Agriculture

Article Title: CRISPR/Cas9-mediated knock-out of SLC15A4 gene involved immune response in bovine rumen epithelial cells1

doi: 10.1016/j.jia.2023.06.016

Figure Lengend Snippet: Fig. 6 Effect of muramyl dipeptide (MDP) on NF-κB and MAPK signaling pathways in wild-type (WT) and SLC15A4-Knockout bovine rumen epithelial cells (BRECs) (KO). The BRECs were treated with 10 μg mL–1 MDP for 6 h. The immunofluorescence for phosphorylation-NF-κB (p-p65) and phosphorylation-ERK1/2 (p-p44) (red) was measured using the nuclear dye 4´,6-diamidino-2-phenylindole (DAPI; blue). Scale bar=100 µm.

Article Snippet: After removing the blocking solution, the phosphorylationERK1/2 (p-p44/42) rabbit mAb (1:1 000; Cell Signaling Technology, Shanghai, China) and phosphorylation-nuclear factor κB (NF-κB) (p-p65) rabbit mAb (1:500; Cell Signaling Technology, Shanghai, China) were added and incubated at 4°C for the duration of the next day.

Techniques: Protein-Protein interactions, Knock-Out, Immunofluorescence, Phospho-proteomics

Journal: Osteoarthritis and Cartilage Open

Article Title: CCL2 induces articular chondrocyte MMP expression through ERK and p38 signaling pathways

doi: 10.1016/j.ocarto.2020.100136

Figure Lengend Snippet: Fig. 2. Effect of CCL2 treatment on ERK, p38 and JNK phosphorylation in normal chondrocytes. Confluent normal human articular chondrocytes were treated with recombinant CCL2 (20ng/ml) for 5, 10 or 15 min. FN-f (1 μM) treatment for 30 min was used as positive control. (A) Cell lysates were subjected to immu- noblotting to detect phospho-ERK, phospho-p38 and phospho-JNK. Immunoblots are representative of n¼4 independent donors. (B) Results of the densitometric analysis showing the effect of CCL2 treatment on the phosphorylation of ERK, p38 and JNK normalized to their respective loading controls. Data are presented as mean values SD. Statistical significance was set at p 0.05 (Asterisks represent 1way ANOVA summary; p values obtained with Tukey’s multiple comparisons are indicated on plots; ns ¼ not significant).

Article Snippet: Antibodies to phospho-ERK (Thr202/Tyr204) (#9101), total-ERK (#9102), phospho-p38 (Thr180/Tyr182) (#9211), total-p38 (#9212), phospho-JNK (Thr183/Tyr185) (#9253), total JNK (#9252) and MMP2 (#4022) were purchased from Cell Signaling Technology.

Techniques: Phospho-proteomics, Recombinant, Positive Control, Western Blot

Journal: Osteoarthritis and Cartilage Open

Article Title: CCL2 induces articular chondrocyte MMP expression through ERK and p38 signaling pathways

doi: 10.1016/j.ocarto.2020.100136

Figure Lengend Snippet: Fig. 3. Effect of CCL2 treatment on ERK and p38 phosphorylation in normal chondrocyte with or without CCR2 inhibition. Confluent normal human articular chondrocytes were treated with recombinant CCL2 (20ng/ml) for 10 min. Where indicated, chondrocytes were pre-incubated with RS504393 (20 μM) for 1h prior to CCL2 treatment. (A) Immunoblotting analyses were performed to detect phospho-ERK and phospho-p38. Immunoblots are representative of n¼4 independent donors. (B) Results of the densitometric analysis showing the effect of CCL2 treatment on the phosphorylation of ERK and p38 with or without CCR2 inhibition normalized to their respecting loading controls. Data are presented as mean values SD. Statistical significance was set at p 0.05 (Asterisks represent 1way ANOVA summary; p values obtained with Tukey’s multiple comparisons are indicated on plots).

Article Snippet: Antibodies to phospho-ERK (Thr202/Tyr204) (#9101), total-ERK (#9102), phospho-p38 (Thr180/Tyr182) (#9211), total-p38 (#9212), phospho-JNK (Thr183/Tyr185) (#9253), total JNK (#9252) and MMP2 (#4022) were purchased from Cell Signaling Technology.

Techniques: Phospho-proteomics, Inhibition, Recombinant, Incubation, Western Blot

Journal: Osteoarthritis and Cartilage Open

Article Title: CCL2 induces articular chondrocyte MMP expression through ERK and p38 signaling pathways

doi: 10.1016/j.ocarto.2020.100136

Figure Lengend Snippet: Fig. 6. Effect of CCL2 treatment on expression of MMPs in OA chondrocytes in presence or absence of ERK and p38 inhibition. Human OA chondrocytes were treated with recombinant CCL2 (20ng/ml) for 10 min. Where indicated, chondrocytes were pre-incubated with ERK inhibitor (U0126, 10 μM) or p38 inhibitor (SB203580, 10 μM), for 1h prior CCL2 treatment. (A) Cell lysates were subjected to immunoblotting to detect phospho-ERK and phospho-p38. Immunoblots are representative of n¼4 independent donors. (B) Results of the densitometric analysis showing the effect of CCL2 treatment on the phosphorylation of ERK and p38 with or without ERK or p38 inhibition normalized to their respective loading controls. (C) Quantitative RT-PCR analyses were performed using the following probes: Mmp3 (n ¼ 6) and Mmp13 (n¼6). Data are presented as mean values SD. Statistical significance was set at p 0.05 (Asterisks represent 1way ANOVA summary; p values obtained with Tukey’s multiple comparisons are indicated on plots; ####p 0.0001).

Article Snippet: Antibodies to phospho-ERK (Thr202/Tyr204) (#9101), total-ERK (#9102), phospho-p38 (Thr180/Tyr182) (#9211), total-p38 (#9212), phospho-JNK (Thr183/Tyr185) (#9253), total JNK (#9252) and MMP2 (#4022) were purchased from Cell Signaling Technology.

Techniques: Expressing, Inhibition, Recombinant, Incubation, Western Blot, Phospho-proteomics, Quantitative RT-PCR