Journal: ACS Omega

Article Title: Mechanisms and Molecular Targets of BuShenHuoXue Formula for Osteoarthritis

doi: 10.1021/acsomega.1c07270

Figure Lengend Snippet: Relative mRNA expression rates of genes in chondrocytes by PCR. (A) SELE. (B) MMP2. (C) COL1. * P < 0.05, ** P < 0.01, *** P < 0.001 versus the IL-1β group.

Article Snippet: The following antibodies and corresponding dilution ratios were used: anti-SELE (bs-1273R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-MMP2 (bs-0412R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-CoL1 (bs-10423R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), and anti-GAPDH (60004-1-lg, Mouse, Proteintech, 120,000, secondary antibody, 1:5000).

Techniques: Expressing

Journal: ACS Omega

Article Title: Mechanisms and Molecular Targets of BuShenHuoXue Formula for Osteoarthritis

doi: 10.1021/acsomega.1c07270

Figure Lengend Snippet: Western blot in five groups of chondrocytes. (A) SELE. (B) MMP2. (C) COL1. (D) Electrophoretic images. ** P < 0.01, *** P < 0.001 versus IL-1β group.

Article Snippet: The following antibodies and corresponding dilution ratios were used: anti-SELE (bs-1273R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-MMP2 (bs-0412R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-CoL1 (bs-10423R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), and anti-GAPDH (60004-1-lg, Mouse, Proteintech, 120,000, secondary antibody, 1:5000).

Techniques: Western Blot

Journal: ACS Omega

Article Title: Mechanisms and Molecular Targets of BuShenHuoXue Formula for Osteoarthritis

doi: 10.1021/acsomega.1c07270

Figure Lengend Snippet: KEGG Enrichment Analysis

Article Snippet: The following antibodies and corresponding dilution ratios were used: anti-SELE (bs-1273R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-MMP2 (bs-0412R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-CoL1 (bs-10423R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), and anti-GAPDH (60004-1-lg, Mouse, Proteintech, 120,000, secondary antibody, 1:5000).

Techniques:

Journal: ACS Omega

Article Title: Mechanisms and Molecular Targets of BuShenHuoXue Formula for Osteoarthritis

doi: 10.1021/acsomega.1c07270

Figure Lengend Snippet: PCR Primer Sequences

Article Snippet: The following antibodies and corresponding dilution ratios were used: anti-SELE (bs-1273R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-MMP2 (bs-0412R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), anti-CoL1 (bs-10423R, Rabbit, Bioss, 1:500, secondary antibody: 1:5000), and anti-GAPDH (60004-1-lg, Mouse, Proteintech, 120,000, secondary antibody, 1:5000).

Techniques: Sequencing

Journal: Molecular Medicine Reports

Article Title: Girdin regulates the migration and invasion of glioma cells via the PI3K-Akt signaling pathway

doi: 10.3892/mmr.2015.4049

Figure Lengend Snippet: Girdin silencing inhibits the expression and activity of MMP-2 and MMP-9. (A and B) Changes in the mRNA levels of MMP-2 and MMP-9 were measured using reverse transcription-quantitative polymerase chain reaction following transfection. The relative mRNA expression levels were calculated using the 2 −ΔΔCt method. (C and D) Following transfection, changes in the protein levels of MMP-2 and MMP-9 were detected using western blot analysis. (E and F) Following transfection, gelatin zymography was performed to detect changes in the activities of MMP-2 and MMP-9. Each experiment was repeated three times. The experimental results are presented as the mean ± standard deviation. ** P<0.01, compared with the NC group. shRNA, short hairpin RNA; NC, negative control.

Article Snippet: Following washing with Tris-buffered saline with 0.05% Tween-20 (TBST), the membranes were incubated with the following primary antibodies at 4°C overnight: Rabbit anti-human polyclonal antibody against girdin (1:500 diluted; cat. no. bs-5150R; Bioss, Beijing, China); rabbit anti-human polyclonal antibody against MMP-2 (1:1,000 diluted; cat. no. WL0657); rabbit anti-human polyclonal antibody against MMP-9 (1:1,000 diluted; cat. no. WL0884); rabbit anti-human polyclonal antibody against P85α (1:1,000 diluted; cat. no. WL0191); rabbit anti-human polyclonal antibody against P110α (1:1,000 diluted; cat. no. WL0339); rabbit anti-human polyclonal antibody against AKT (1:1,000 diluted; cat. no. WL0003); rabbit anti-human polyclonal antibody against p-AKT (1:1,000 diluted; cat. no. WLP001).

Techniques: Expressing, Activity Assay, Real-time Polymerase Chain Reaction, Transfection, Western Blot, Zymography, Standard Deviation, shRNA, Negative Control

Journal: Epigenetics

Article Title: Methylation of TMEM176A is an independent prognostic marker and is involved in human colorectal cancer development

doi: 10.1080/15592294.2017.1341027

Figure Lengend Snippet: TMEM176A inhibits cell proliferation, colony formation, migration, and invasion, and induces apoptosis in CRC cells. (A) Effect of TMEM176A on cell proliferation measured by the MTT assay. The experiment was repeated 3 times. *: P<0.05. NC: siRNA negative control. (B) Representative results of colony formation in TMEM176A unexpressing- and TMEM176A re-expressing-RKO and TMEM176A unexpressing- and TMEM176A re-expressing-DLD1 cells. The experiment was repeated 3 times. *: P<0.05. (C) Percentage of apoptotic cells before and after restoration of TMEM176A expression in RKO and DLD1 cells before and after knockdown of TMEM176A in SW620 cells. The experiment was repeated 3 times. *: P<0.05. (D) Transwell assay showing migration and invasion results before and after restoration of TMEM176A expression in RKO and DLD1 cells and before and after knockdown of TMEM176A in SW620 cells (200 ×). The experiment was repeated 3 times. (E) Expression levels of TMEM176A, cleaved caspase-3, MMP-2, MMP-7, and MMP-9 were detected by Western blot before and after restoration of TMEM176A expression in RKO and DLD1 cells and before and after knockdown of TMEM176A in SW620 cells.

Article Snippet: Primary antibodies were as follows: anti-TMEM176A (Sigma, St. Louis, MO), anti-cleaved caspase-3 (Cell Signaling Technology), anti-MMP2 (Bioworld Tech., MN, USA), anti-MMP7 (Bioworld Tech.), anti-MMP9 (Bioworld Tech.) and anti-β-actin (Beyotime Biotech, China).

Techniques: Migration, MTT Assay, Negative Control, Expressing, Transwell Assay, Western Blot

Journal: Epigenetics

Article Title: Methylation of TMEM176A is an independent prognostic marker and is involved in human colorectal cancer development

doi: 10.1080/15592294.2017.1341027

Figure Lengend Snippet: TMEM176A suppresses CRC cell xenograft tumor growth in mice. (A) Representative tumors from TMEM176A unexpressing- and TMEM176A re-expressing-DLD1 cell xenografts. (B) Tumor growth curves of TMEM176A unexpressing- and TMEM176A re-expressing-DLD1 cells. *: P<0.05. (C) Tumor weights in nude mice at the 31st day after inoculation of TMEM176A for TMEM176A unexpressing- and TMEM176A re-expressing-DLD1 cells. Bars: mean of 5 mice. *: P<0.05. (D) Images of hematoxylin and eosin staining show tumors from TMEM176A unexpressing- and TMEM176A re-expressing-DLD1 xenograft mice. IHC staining reveals the expression levels of TMEM176A, MMP2, MMP7, MMP9, and cleaved caspase-3 in TMEM176A unexpressing- and TMEM176A re-expressing-DLD1 cell xenografts (200 ×).

Article Snippet: Primary antibodies were as follows: anti-TMEM176A (Sigma, St. Louis, MO), anti-cleaved caspase-3 (Cell Signaling Technology), anti-MMP2 (Bioworld Tech., MN, USA), anti-MMP7 (Bioworld Tech.), anti-MMP9 (Bioworld Tech.) and anti-β-actin (Beyotime Biotech, China).

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Cancer biology & therapy

Article Title: The involvement of MALAT1-ALKBH5 signaling axis into proliferation and metastasis of human papillomavirus-positive cervical cancer.

doi: 10.1080/15384047.2023.2249174

Figure Lengend Snippet: Figure 6. Effect of the MALAT1-ALKBH5 axis on MMP2 and MMP9 expression. (a and b) HeLa and CaSki cells were, respectively, transfected with NC and two siRNAs against MALAT1 or ALKBH5. The expression levels of MMP2 and MMP9 was determined by RT-qPCR. (c) The cell lysates were subjected to western blot analysis of MMP2 and MMP9 after knockdown of MALAT1 in HeLa and CaSki cells using GAPDH as loading control. (d) Both HeLa and CaSki cells were transfected with NC and siRNAs against MALAT1 with co-transfection of the ALKBH5 expression vector (pcDNA3.1-ALKBH5), with empty vector (pcDNA3.1) serving as the control. Cell RNA was isolated and subsequently reverse transcribed into cDNA, which were subjected to RT-qPCR for determination of MMP2 and MMP9 expression using specific primers. NC, negative control.

Article Snippet: The antibody information was as follows: Primary antibody: anti-ALKBH5 antibody (1:1000, Cell Signaling Technology, Inc.), anti-MMP2 antibody (1:1000, Elabscience Inc.), anti-MMP9 antibody (1:1000, BOSTER Biological Technology Co. Ltd.), and anti-GAPDH antibody (1:5000, Beyotime Institute of Biotechnology); and secondary antibody: HRP conjugated anti-rabbit or -mouse secondary antibody (1:5000, MultiSciences Biotech Co., Ltd.).

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Knockdown, Control, Cotransfection, Plasmid Preparation, Isolation, Reverse Transcription, Negative Control