primary antibodies against mmp 1  (Servicebio Inc)

Journal: Molecular Medicine Reports

Article Title: Tributyltin chloride induces chondrocyte damage through the activation of NLRP3‑mediated inflammation and pyroptosis

doi: 10.3892/mmr.2024.13247

Figure Lengend Snippet: TBTC induces chondrocyte damage in rats. (A) Cell Counting Kit-8 results showed that TBTC reduced rat chondrocyte viability in a concentration-dependent manner. (B) TBTC significantly increased LDH leakage from rat chondrocytes. (C) Reverse transcription-quantitative PCR results revealed that TBTC elevated NLRP3 mRNA expression levels. (D) Western blot analysis results revealed that TBTC upregulated the protein expression levels of NLRP3, IL-1β, IL-18, MMP-1 and MMP-13 in rat chondrocytes. *P<0.05, **P<0.01, ***P<0.001 vs. Con. Con, control; IL, interleukin; LDH, lactate dehydrogenase; MMP, matrix metalloproteinase; NLRP3, NLR family pyrin domain containing 3; TBTC, tributyltin chloride.

Article Snippet: Subsequently, primary antibodies against β-actin (1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.), NLRP3 (1:1,000; cat. no. ab263899; Abcam), interleukin (IL)-1β (1:1,000; cat. no. ab315084; Abcam), IL-18 (1:1,000; cat. no. ab223293; Abcam), matrix metalloproteinase (MMP)-1 (1:500; cat. no. 70R-50066; AmyJet Scientific, Inc.), MMP-13 (1:1,000; cat. no. ab219620; Abcam), caspase-1 (1:1,000; cat. nos. ab286125 for rat or ab138483 for mouse; Abcam), PYD and CARD domain containing (ASC; 1:1,000; cat. no. ab307560; Abcam) and gasdermin D (GSDMD; 1:1,000; cat. no. ab219800; Abcam) were incubated with the PVDF membranes overnight at 4°C.

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

Journal: Molecular Medicine Reports

Article Title: Tributyltin chloride induces chondrocyte damage through the activation of NLRP3‑mediated inflammation and pyroptosis

doi: 10.3892/mmr.2024.13247

Figure Lengend Snippet: TBTC enhances the NLRP3-mediated inflammatory response and cellular pyroptosis in mouse OA. (A) TBTC increased the protein expression levels of NLRP3, IL-1β, IL-18, MMP-1 and MMP-13. (B) TBTC increased the expression levels of the pyroptosis-related proteins caspase-1, ASC and GSDMD in mouse osteoarticular tissues. *P<0.05, **P<0.01, ***P<0.001 vs. Con; #P<0.05, ##P<0.01, ###P<0.001 vs. OA. Con. ASC, PYD and CARD domain containing; Con, control; GSDMD, gasdermin D; IL, interleukin; MMP, matrix metalloproteinase; OA, osteoarthritis; TBTC, tributyltin chloride.

Article Snippet: Subsequently, primary antibodies against β-actin (1:1,000; cat. no. 4970; Cell Signaling Technology, Inc.), NLRP3 (1:1,000; cat. no. ab263899; Abcam), interleukin (IL)-1β (1:1,000; cat. no. ab315084; Abcam), IL-18 (1:1,000; cat. no. ab223293; Abcam), matrix metalloproteinase (MMP)-1 (1:500; cat. no. 70R-50066; AmyJet Scientific, Inc.), MMP-13 (1:1,000; cat. no. ab219620; Abcam), caspase-1 (1:1,000; cat. nos. ab286125 for rat or ab138483 for mouse; Abcam), PYD and CARD domain containing (ASC; 1:1,000; cat. no. ab307560; Abcam) and gasdermin D (GSDMD; 1:1,000; cat. no. ab219800; Abcam) were incubated with the PVDF membranes overnight at 4°C.

Techniques: Expressing

Journal: Journal of Microbiology and Biotechnology

Article Title: 1-Kestose Blocks UVB-Induced Skin Inflammation and Promotes Type I Procollagen Synthesis via Regulating MAPK/AP-1, NF-κB and TGF-β/Smad Pathway

doi: 10.4014/jmb.2311.11020

Figure Lengend Snippet: ( A, B ) Level of expressed MMP-1 was detected by western blotting 12 h after UVB irradiation. Band intensity was quantified by ImageJ and normalized to β-actin. ( C ) Dual-luciferase reporter assay was evaluated to measure the MMP-1 promoter activity in the HaCaT cells. The mRNA expression of ( D ) MMP-1, ( E ) MMP-3, ( F ) MMP-9, ( G ) COL1A1 and ( H ) COL2A1 was measured using RT-PCR, followed by treatment with 1-kestose (4, 6, and 10 mM) for 12 h. All data are expressed as mean ± SD ( n = 3). # p < .05, ## p < .01, ### p < .001, and #### p < .0001 vs. blank group, * p < .05, ** p < .01, *** p < .001, and **** p < .0001 vs control group.

Article Snippet: Primary antibodies against MMP-1 (#54376), ERK (#4695), JNK (#9252), p38 (#8690), p-ERK (#4370), p-JNK (#4671), p-p38 (#4511), c-Fos, (#2250), c-Jun (#9165), p-c-Fos (#5348), p-c-Jun (#3270), TGF-β1 (#3711), Smad2/3 (#8685), p-Smad2/3 (#8828), and β-actin (#4970), as well as horseradish peroxidase (HRP) goat anti-rabbit IgG (#2895) were purchased from Cell Signaling Technology (USA).

Techniques: Western Blot, Irradiation, Luciferase, Reporter Assay, Activity Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Cell Death Discovery

Article Title: IL-8 activates fibroblasts to promote the invasion of HNSCC cells via STAT3-MMP1

doi: 10.1038/s41420-024-01833-7

Figure Lengend Snippet: A MMP1 expression profile across all tumor samples and paired normal tissues in TCGA. Compare with other primary tumor samples, MMP1 expression level was significantly upregulated in HNSCC samples. B Kaplan–Meier analysis of overall survival. Compared with patients with low MMP1 expression, those with high MMP1 expression had a significantly lower overall survival rate. C , D MMP1 expression in HNSCC samples and paired normal tissues (tumor/normal) from 53 patients were examined by qRT-PCR (brown for MMP1 down-regulation in tumor tissues compared to normal tissues while orange for up-regulation in tumor tissues) and those from another 92 patients were examined by immunohistochemistry staining. (−: negative; +: positive; ++: strongly positive).

Article Snippet: Primary antibodies against MMP1 (#54376; CST), Phospho-STAT3 Tyr705 (#9145; CST), STAT3 (#12640; CST), GAPDH (HRP-60004; Proteintech, China), Lamin A/C (10298-1-AP; Proteintech, China), Alpha Tubulin (HRP-66031; Proteintech, China) in 3% bovine serum albumin and secondary antibody (Mouse BA1050; Rabbit BA1054; Boster, China) were used.

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry, Staining

Journal: Cell Death Discovery

Article Title: IL-8 activates fibroblasts to promote the invasion of HNSCC cells via STAT3-MMP1

doi: 10.1038/s41420-024-01833-7

Figure Lengend Snippet: A , B The transcriptional and translational levels of MMP1 were detected by qRT-PCR and western blotting in HNSCC cell line (HN6). MMP1 was downregulated in both. The migration abilities of HNSCC cells with MMP1 knockdown were detected using Transwell assays ( C ) and wound-healing assays ( D ) respectively. The migration abilities of MMP-1-knockdown HNSCC cells treated with rhMMP1 (at 24 h, 20 ng/ml) were detected using Transwell assays ( E ) and wound-healing assays ( F ) respectively; Data are presented as the mean ± SD from three independent experiments (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Primary antibodies against MMP1 (#54376; CST), Phospho-STAT3 Tyr705 (#9145; CST), STAT3 (#12640; CST), GAPDH (HRP-60004; Proteintech, China), Lamin A/C (10298-1-AP; Proteintech, China), Alpha Tubulin (HRP-66031; Proteintech, China) in 3% bovine serum albumin and secondary antibody (Mouse BA1050; Rabbit BA1054; Boster, China) were used.

Techniques: Quantitative RT-PCR, Western Blot, Migration, Knockdown

Journal: Cell Death Discovery

Article Title: IL-8 activates fibroblasts to promote the invasion of HNSCC cells via STAT3-MMP1

doi: 10.1038/s41420-024-01833-7

Figure Lengend Snippet: A MMP1 expression among identified cell types in single-cell RNA-seq dataset (GSE103322). B The immuno-colocalization of MMP1 (red) and ACTA2 (green) was detected in HNSCC samples by fluorescence microscope, and was represented as the overlapping fluorescence for each channel, where the combined pixels were yellow. C The expression of MMP1 protein levels in different HNSCC cell lines (LN4, Cal27, HN6, HN30) versus NFs. D The images of invaded transwell assay for HN6 cells (24 h) showed that each group of NF conditioned mediums promoted the invasive abilities of cancer cells compare to blank control (DMEM with 10% FBS). Each group of fibroblast was starved overnight and conditioned mediums were collected. E The downregulation of MMP1 protein expression in NF3 was demonstrated by western blotting. F Indirect co-culture invaded transwell assays were performed. Each group of NF3 fibroblasts were transfected with siRNA and cultured on the 24-well plates overnight, and the conditioned medium of each was collected for the lower layer of the matrigel-invasion chambers. The results showed fewer invasive tumor cells were detected compared to the siNC group. The chemotactic effect of NF3 on HN6 (24 h) and CAL27 (48 h) were attenuated with downregulation of MMP1.

Article Snippet: Primary antibodies against MMP1 (#54376; CST), Phospho-STAT3 Tyr705 (#9145; CST), STAT3 (#12640; CST), GAPDH (HRP-60004; Proteintech, China), Lamin A/C (10298-1-AP; Proteintech, China), Alpha Tubulin (HRP-66031; Proteintech, China) in 3% bovine serum albumin and secondary antibody (Mouse BA1050; Rabbit BA1054; Boster, China) were used.

Techniques: Expressing, RNA Sequencing, Fluorescence, Microscopy, Transwell Assay, Control, Western Blot, Co-Culture Assay, Transfection, Cell Culture

Journal: Cell Death Discovery

Article Title: IL-8 activates fibroblasts to promote the invasion of HNSCC cells via STAT3-MMP1

doi: 10.1038/s41420-024-01833-7

Figure Lengend Snippet: A , B MMP1 proteins in NFs were detected at indicated time after CM treatment by western blotting assays. The expression of MMP1 protein levels in NFs was upregulated after 12-h HN30-CM treatment. C Heatmap of differential expression of selected proteins in culture supernatants (HN30-CM compared with 293T-CM). IL6 and IL8 were the higher components. D IL6, IL8 (CXCL8), STAT3 and MMP1 are relevant according to STRING network clusters. E HN30-CM induces STAT3 phosphorylation in NFs within 2 h, and upregulates the expression of MMP1 after 12 h. F HN30 conditioned medium increased phosphorylated STAT3 protein levels in nuclear fractions. G ChIP assay was used to assess the binding of STAT3 to the promoters of the indicated genes in NF3 or HN30-CM mediated (1 h for treatment) NF3 with anti-STAT3 antibodies, followed by qPCR analysis.

Article Snippet: Primary antibodies against MMP1 (#54376; CST), Phospho-STAT3 Tyr705 (#9145; CST), STAT3 (#12640; CST), GAPDH (HRP-60004; Proteintech, China), Lamin A/C (10298-1-AP; Proteintech, China), Alpha Tubulin (HRP-66031; Proteintech, China) in 3% bovine serum albumin and secondary antibody (Mouse BA1050; Rabbit BA1054; Boster, China) were used.

Techniques: Western Blot, Expressing, Quantitative Proteomics, Phospho-proteomics, Binding Assay

Journal: Cell Death Discovery

Article Title: IL-8 activates fibroblasts to promote the invasion of HNSCC cells via STAT3-MMP1

doi: 10.1038/s41420-024-01833-7

Figure Lengend Snippet: A – C Phospho-STAT3, STAT3 proteins were detected 0.5 h after treatment by western blotting assays. After NF3 cells were cultured in basal medium and starved overnight, IgG, anti-IL6R, anti-IL6ST or anti-IL6 and anti-IL8 (100 ng/ml) were added before treating with HN30-CM and IL6 recombinant protein (10 ng/ml) or IL8 recombinant protein (10-50 ng/ml) for 0.5 hours. D MMP1 proteins were detected 12 hours after HN30-CM or together with IL6 or IL8 antibody (100 ng/ml) treatment in NF3 cells by western blotting assays. E The images of indirect co-culture invaded transwell assays. Invasion of HN6 and CAL27 cells cultured with NF3 (control without treatment), NF3 pretreated with HN30-CM (positive), NF3 pretreated together with IL6 antibody and HN30-CM, NF3 pretreated together with IL8 antibody and HN30-CM.

Article Snippet: Primary antibodies against MMP1 (#54376; CST), Phospho-STAT3 Tyr705 (#9145; CST), STAT3 (#12640; CST), GAPDH (HRP-60004; Proteintech, China), Lamin A/C (10298-1-AP; Proteintech, China), Alpha Tubulin (HRP-66031; Proteintech, China) in 3% bovine serum albumin and secondary antibody (Mouse BA1050; Rabbit BA1054; Boster, China) were used.

Techniques: Western Blot, Cell Culture, Recombinant, Co-Culture Assay, Control

Journal: Cell Death Discovery

Article Title: IL-8 activates fibroblasts to promote the invasion of HNSCC cells via STAT3-MMP1

doi: 10.1038/s41420-024-01833-7

Figure Lengend Snippet: A , B Phospho-STAT3, STAT3 and MMP1 proteins are detected in indicated time by western blotting assay. NF3 cells were serum-starved and A treated with CM from HN30 and TPCA (inhibitor of STAT3 signaling pathway) for the indicated time or B treated with TPCA and CM from HN30 with indicated contents for 12 hours. C Invasion of HN6 cells cultured with NF3, NF3 pretreated with CM from HN30 cells or NF3 pretreated with TPCA and CM from HN30 cells.

Article Snippet: Primary antibodies against MMP1 (#54376; CST), Phospho-STAT3 Tyr705 (#9145; CST), STAT3 (#12640; CST), GAPDH (HRP-60004; Proteintech, China), Lamin A/C (10298-1-AP; Proteintech, China), Alpha Tubulin (HRP-66031; Proteintech, China) in 3% bovine serum albumin and secondary antibody (Mouse BA1050; Rabbit BA1054; Boster, China) were used.

Techniques: Western Blot, Cell Culture

Journal: Cell Death Discovery

Article Title: IL-8 activates fibroblasts to promote the invasion of HNSCC cells via STAT3-MMP1

doi: 10.1038/s41420-024-01833-7

Figure Lengend Snippet: A , B Representative H&E images and graphical analysis of frequency of micrometastatic foci and lymph node metastasis. After 106 LN4 OSCC cells were mixed with 106 NF-SCR-sh or NF-MMP1-sh and injected into the tongue of 5-week-old nude mice. n = 10/group. **, P < 0.01; ***, P < 0.005; P = two tailed t test or Fisher’s exact test. C Proposed working model for the prometastatic roles of MMP1 and STAT3 in fibroblasts. Cytokine IL-8 is secreted by tumor cells, which activates fibroblasts around. Thus, in response to IL-8, STAT3 in fibroblasts are phosphorylated and translocated into the nucleus. As a result, the STAT3-dependent MMP1 gene encoding secreted proteins is induced. Those in turn exert pro-migratory and pro-invasive functions on tumor cells.

Article Snippet: Primary antibodies against MMP1 (#54376; CST), Phospho-STAT3 Tyr705 (#9145; CST), STAT3 (#12640; CST), GAPDH (HRP-60004; Proteintech, China), Lamin A/C (10298-1-AP; Proteintech, China), Alpha Tubulin (HRP-66031; Proteintech, China) in 3% bovine serum albumin and secondary antibody (Mouse BA1050; Rabbit BA1054; Boster, China) were used.

Techniques: Injection, Two Tailed Test