Journal: Science advances

Article Title: Selective promotion of sensory innervation-mediated immunoregulation for tissue repair.

doi: 10.1126/sciadv.ads9581

Figure Lengend Snippet: Fig. 8. Sema3A mediates XIAP expression to promote the M2 polarization of macrophages. (A) Schematic representation of macrophage conditioned culture. The super- natant of DRGC culture medium was collected and mixed with the conventional culture medium of macrophages in equal proportion to obtain the CM. (B) Immunofluores- cence staining and (C) quantification of CD206 (M2 marker) and iNOS (M1 marker) expression in macrophages. Scale bar, 30 μm. (D) ELISAs of anti-inflammatory cytokine IL-10 and pro-inflammatory cytokines TNF-α and IL-1β. (E) Immunofluorescence staining and (F) the corresponding quantification of CD206 and iNOS expression in macrophages treated with CM from Sema3A-specific knockdown DRGCs and rSema3A. Scale bars, 30 μm. (G) ELISAs of IL-10, TNF-α, and IL-1β expression after culturing with different CMs. (H) Volcano plot of the differential gene expression in macrophages after Sema3A transfection. (I) Heatmap of the mRNA transcription profiles of the top 15 up-/down- regulated genes. (J) Venn diagram indicating the number of genes overlapping with those collected from the GSEA database (inflammation-related gene sets containing 303 different genes) and the top 15 up-/down-regulated genes screened using RNA-seq. Two genes (Xiap and Mmp13) were overlapping. (K) Representative Western blotting images of XIAP and MMP13 in macrophages after culturing with Sema3A. (L and M) Immunofluorescence images and the corresponding fluorescence intensity of CD206 and iNOS expression in macrophages after shXIAP transfection. Scale bar, 30 μm. (N) ELISAs of IL-10, TNF-α, and IL-1β after shXIAP transfection. (O and P) Immunofluorescence images and the corresponding quantitative analysis of CD206 and iNOS in XIAP-overexpressing macrophages transfected with pcDNA3.1-XIAP. Scale bar, 30 μm. (Q) ELISAs of IL-10, TNF-α, and IL-1β in macrophages overexpressing XIAP. IAN, inferior alveolar nerve; rSema3A, recombinant Sema3A. **P < 0.01; ***P < 0.001; NS, not significant.

Article Snippet: The antibodies used in this part of experiments include Sema3A (1:1000, 27836- 1- AP, Proteintech), MMP13 (1:2000, 181651- AP, Proteintech), XIAP (1:2000, A20846, Abclonal), PAX6 (1:500, A19099, Abclonal), β- actin (1:100000, AC026, Abclonal), Flag- Tag (80010- 1- RR, Proteintech), Myc- Tag (60003- 2- Ig, Proteintech), HA- Tag (#2367, Cell Signaling Technology, USA), and anti- rabbit/mouse- HRP (BA1056, Boster, USA).

Techniques: Expressing, Staining, Marker, Immunofluorescence, Knockdown, Gene Expression, Transfection, RNA Sequencing, Western Blot, Fluorescence, Recombinant

Journal: Drug Design, Development and Therapy

Article Title: S-nitroso-N-acetylcysteine attenuates liver fibrosis in experimental nonalcoholic steatohepatitis

doi: 10.2147/DDDT.S43930

Figure Lengend Snippet: Fold change in matrix metalloproteinase (MMP)-13 and -9, and tissue inhibitor of metalloproteinases (TIMP)-1 and -2 mRNA expression in the nonalcoholic steatohepatitis groups against S-nitroso-N-acetylcysteine groups. Abbreviations: NASH, nonalcoholic steatohepatitis; SNAC, S-nitroso-N-acetylcysteine.

Article Snippet: All primers and probes were purchased from Applied Biosystems (Assay ID: β-actin: 4331348; collagen-1a: Rn00801649_q1; TGFβ-1: Rn00572010_m1; MMP-2: Rn01538167_m1; TIMP-1: Rn00587558_m1 and MMP-13: Rn01448191_g1).

Techniques: Expressing

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: Effect of CCL2 treatment on the expression of Ccr2 receptor, Mmp1 , Mmp3 , Mmp13 and Timp1 in normal chondrocytes. RNA was isolated from confluent normal human articular chondrocytes treated with recombinant CCL2 (20ng/ml) for 6, 24 or 72 ​h. Quantitative RT-PCR analyses was performed using the following probes: Ccr2 ( n = 6 ), Mmp1 ( n ​= ​6), Mmp3 (6h, n ​= ​9; 24–72h, n ​= ​6), Mmp13 (6h, n ​= ​10; 24h, n ​= ​7; 72h, n ​= ​6), and Timp1 ( n = 6 ). Data are presented as mean values ​± ​SD. Statistical significance was set at p ​≤ ​0.05 (Unpaired t -test).

Article Snippet: Target primer/probes included were MMP1 (Hs00899658_m1); MMP3 (Hs00968305_m1); MMP13 (Hs00942584_m1); CCR2 (Hs00704702_s1); TIMP1 (Hs01092512_g1); COMP (Hs00164359_m1); COL2A1 (Hs00264051_m1); ADAMTSL4 (Hs01120103_g1); ADAMTSL5 (Hs01358655_g1).

Techniques: Expressing, Isolation, Recombinant, Quantitative RT-PCR

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: Effect of CCL2 treatment on expression of MMPs in the presence or absence of ERK and p38 inhibition in normal chondrocytes. Confluent normal human articular 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 to CCL2 treatment. ( A ) Cell lysates were subjected to immunoblotting to detect phospho-ERK and phospho-p38. Immunoblots are representative of n = 5 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: Mmp1 ( n ​= ​6), Mmp3 (6h, n ​= ​9; 24–72h, n ​= ​6), Mmp13 (6h, n ​= ​10; 24h, n ​= ​7; 72h, n ​= ​6), and Timp1 ( n = 6 ). 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; ns ​= ​not significant).

Article Snippet: Target primer/probes included were MMP1 (Hs00899658_m1); MMP3 (Hs00968305_m1); MMP13 (Hs00942584_m1); CCR2 (Hs00704702_s1); TIMP1 (Hs01092512_g1); COMP (Hs00164359_m1); COL2A1 (Hs00264051_m1); ADAMTSL4 (Hs01120103_g1); ADAMTSL5 (Hs01358655_g1).

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

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: Effect of CCL2 treatment on the expression of Ccr2 , Mmp1 , Mmp3 , Mmp13 and Timp1 in OA chondrocytes. RNA was isolated from confluent human articular OA chondrocytes treated with recombinant CCL2 (20ng/ml) for 6, 24 or 72 ​h. Quantitative RT-PCR analyses were performed using the following probes: Ccr2 ( n = 6 ), Mmp1 ( n ​= ​6), Mmp3 ( n ​= ​6), Mmp13 ( n = 6 ), and Timp1 ( n = 6 ). Data are presented as mean values ​± ​SD. Statistical significance was set at p ​≤ ​0.05 (Unpaired t -test; ns ​= ​not significant).

Article Snippet: Target primer/probes included were MMP1 (Hs00899658_m1); MMP3 (Hs00968305_m1); MMP13 (Hs00942584_m1); CCR2 (Hs00704702_s1); TIMP1 (Hs01092512_g1); COMP (Hs00164359_m1); COL2A1 (Hs00264051_m1); ADAMTSL4 (Hs01120103_g1); ADAMTSL5 (Hs01358655_g1).

Techniques: Expressing, Isolation, Recombinant, Quantitative RT-PCR

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: 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 n = 4 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 ​= ​6) and Mmp13 ( n = 6 ). 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: Target primer/probes included were MMP1 (Hs00899658_m1); MMP3 (Hs00968305_m1); MMP13 (Hs00942584_m1); CCR2 (Hs00704702_s1); TIMP1 (Hs01092512_g1); COMP (Hs00164359_m1); COL2A1 (Hs00264051_m1); ADAMTSL4 (Hs01120103_g1); ADAMTSL5 (Hs01358655_g1).

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

Journal: Cell Death & Disease

Article Title: Impact of AMPK on cervical carcinoma progression and metastasis

doi: 10.1038/s41419-023-05583-9

Figure Lengend Snippet: Taq-Man qRT-PCR probes list.

Article Snippet: MMP-1 , Hs00899658_m1 , MMP-13 , Hs00233992_m1.

Techniques: