Journal: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

Article Title: Association of the α 2 δ 1 Subunit with Ca v 3.2 Enhances Membrane Expression and Regulates Mechanically Induced ATP Release in MLO-Y4 Osteocytes

doi: 10.1002/jbmr.437

Figure Lengend Snippet: RT-PCR primers used for detection of VSCC subunits and osteocyte markers

Article Snippet: The rabbit anti-β 3 antibody was purchased from Alomone Research Laboratories (Jerusalem, Israel).

Techniques: Sequencing

Journal: International immunopharmacology

Article Title: Inhibiting the SARM1-NAD + axis reduces oxidative stress-induced damage to retinal and nerve cells.

doi: 10.1016/j.intimp.2024.112193

Figure Lengend Snippet: Fig. 6. DSRM ameliorated oxidative stress-induced neurite injury in N2a cells. (A-C) GOx inhibited N2a cell proliferation and neurite growth in the RA-induced N2a cell differentiation model (magnification 400 × ); (D-E) Immunocytochemistry demonstrated that 10 μM DSRM pre-treatment for 2 h enhanced β3-tubulin expression induced by 7 mU/mL GOx for 12 h in N2a cells (scale bar: 50 μm). N ≥3, **p < 0.01, ****p < 0.0001.

Article Snippet: After culturing and treating the cells in 24-well plates and fixing and permeabilizing them with methanol at -20 ◦C for 15 min, the cells were then washed 3 times with PBS, and then incubated in containment buffer (5 % donkey serum for containment) for 20 min, and then incubated with anti-GSDMD primary antibody (1:500, Abclonal, Wuhan, China, Cat#A20197), anti-β3-tubulin primary antibody (1:1000, CST, USA, Cat#5568S) was incubated at 4 ◦C overnight.

Techniques: Cell Differentiation, Immunocytochemistry, Expressing

Journal: Frontiers in Molecular Neuroscience

Article Title: LRRK2 Regulates Voltage-Gated Calcium Channel Function

doi: 10.3389/fnmol.2016.00035

Figure Lengend Snippet: Biophysical properties of HEK293 cells recorded in different experimental conditions .

Article Snippet: The primary antibodies used were: rabbit anti-LRRK2 at 1:500 (MJFF2, c41-2 Abcam), rabbit anti-calcium channel β3 subunit 1:200 (Sigma), rabbit anti-calcium channel α1a subunit 1:200 (Sigma), mouse anti-actin 1:2000, mouse anti-myc 1:1000 (Sigma), rabbit anti-S6 ribosomal protein (S6RP) 1:1000 (Cell Signalling) applied in blocking buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 0.1% Tween 20) and 5% nonfat dry milk, overnight at 4°C.

Techniques: Transfection

Journal: Frontiers in Molecular Neuroscience

Article Title: LRRK2 Regulates Voltage-Gated Calcium Channel Function

doi: 10.3389/fnmol.2016.00035

Figure Lengend Snippet: LRRK2 interacts with the Ca V 2.1 channel. (A) Mouse forebrain specimens obtained from littermate wt and heterozygous BAC hG2019S (GS) mice were cross-linked and then processed for immunoprecipitation with rat anti LRRK2 antibodies. Eluted proteins were resolved by SDS-PAGE and detected with anti LRRK2, anti Ca V 2.1 β3 subunit antibodies, anti Ca V 2.1 α1a and anti S6 ribosomal protein (S6RP). (B,C) The graphs report the yield of protein recovered upon LRRK2 immunoprecipitation expressed as a percentage of relative input (B) and as amount of pulled Ca V 2.1 β3 subunit relative to the amount of pulled LRRK2 (C) . Bars represent mean ± SEM ( n = 4) where * indicates p < 0.05; unpaired t -test.

Article Snippet: The primary antibodies used were: rabbit anti-LRRK2 at 1:500 (MJFF2, c41-2 Abcam), rabbit anti-calcium channel β3 subunit 1:200 (Sigma), rabbit anti-calcium channel α1a subunit 1:200 (Sigma), mouse anti-actin 1:2000, mouse anti-myc 1:1000 (Sigma), rabbit anti-S6 ribosomal protein (S6RP) 1:1000 (Cell Signalling) applied in blocking buffer (20 mM Tris, pH 7.4, 150 mM NaCl, 0.1% Tween 20) and 5% nonfat dry milk, overnight at 4°C.

Techniques: Immunoprecipitation, SDS Page

Journal: International Journal of Molecular Sciences

Article Title: αvβ3 Integrin as a Link between the Development of Fibrosis and Thyroid Hormones in Systemic Sclerosis

doi: 10.3390/ijms24108927

Figure Lengend Snippet: List of antibodies.

Article Snippet: Integrin β3 , Rabbit , Polyclonal Ab , Santa Cruz Biotechnology , SC14009 , 1:200 , BSA 5%.

Techniques: Hybridization

Journal: Journal of biomedical science

Article Title: Periostin promotes ovarian cancer metastasis by enhancing M2 macrophages and cancer-associated fibroblasts via integrin-mediated NF-κB and TGF-β2 signaling.

doi: 10.1186/s12929-022-00888-x

Figure Lengend Snippet: Fig. 2 POSTN regulates the ERK/NF-κB axis to modulate ovarian cancer cell migration and invasion. A Western blot analysis for detecting phosphoproteins and proteins as indicated in SKOV3 cells transfected with POSTN or in SKOV-I6 cells infected with lentiviral vectors encoding shPOSTN or a scrambled control. B PLA images show protein–protein interactions between POSTN and integrin β3 or integrin β5 in SKOV-I6 cells. The scale bar represents 25 μm. C Western blot analysis for detecting phosphoproteins and proteins as indicated in SKOV3 cells treated with recombinant POSTN protein (100 ng/mL), in combination with anti-αvβ3 (10 μg/mL) or anti-αvβ5 (10 μg/mL) neutralizing antibody as indicated. D Western blot analysis for measuring proteins and phosphoprotein as indicated in SKOV-I6 cells transfected with siITGB5 or control oligonucleotides. E Western blot analysis for detecting p-ERK and ERK in SKOV-I6 cells treated with antibody against POSTN or IgG. F, G Migration and invasion assays of SKOV3/POSTN or SKOV-I6 cells treated with a specific POSTN monoclonal antibody (2 μg/mL) or IgG. **p < 0.01; *** p < 0.001

Article Snippet: Primary antibodies anti-POSTN antibody (SC-46655, Santa Cruz Biotechnology, Dallas, TX, USA), anti-integrin β3 antibody (#13166, Cell Signaling Technology, Danvers, MA, USA) and anti-Integrin β5 antibody (#3629; Cell Signaling Technology) were diluted 1:100 in buffer and added to samples for overnight at 4 °C.

Techniques: Migration, Western Blot, Transfection, Infection, Control, Protein-Protein interactions, Recombinant

Journal: Journal of biomedical science

Article Title: Periostin promotes ovarian cancer metastasis by enhancing M2 macrophages and cancer-associated fibroblasts via integrin-mediated NF-κB and TGF-β2 signaling.

doi: 10.1186/s12929-022-00888-x

Figure Lengend Snippet: Fig. 4 POSTN increases monocytes migration and promotes M2 macrophages polarization. A THP-1 cell migration ability was determined by in vitro migration assay upon addition of the control SKOV3/pcDNA4 or SKOV3/POSTN conditioned medium with or without specific neutralizing antibody pre-treatment in the bottom well. The αvβ3 or αvβ5 integrin neutralizing antibody, as indicated, was added before collecting the conditioned medium. *p < 0.01, ***p < 0.001. B Flow cytometry analysis of THP-1 cells treated with the vector control or POSTN-overexpressing conditioned medium to measure CD68-positive, CD206-positive or CD80-positive populations (left, middle and right, respectively). **p < 0.01, ***p < 0.001. C Flow cytometry analysis of THP-1 cells treated with conditioned medium from the non-specific (N/S) shRNA control or shPOSTN SKOV-I6 cells to measure CD68-positive, CD206-positive or CD80-positive populations (left, middle and right, respectively). *p < 0.05, **p < 0.01. D, E qRT-PCR analysis of THP-1 cells co-cultured with SKOV3 cells transfected with a control or POSTN-overexpressing vector for 5 days to measure M2- or M1-associated cytokines expression change (D, E, respectively). The mRNA expression levels were compared based on the relative fold-change of each gene and the associated p values were calculated using a paired t test. *p < 0.05, **p < 0.01

Article Snippet: Primary antibodies anti-POSTN antibody (SC-46655, Santa Cruz Biotechnology, Dallas, TX, USA), anti-integrin β3 antibody (#13166, Cell Signaling Technology, Danvers, MA, USA) and anti-Integrin β5 antibody (#3629; Cell Signaling Technology) were diluted 1:100 in buffer and added to samples for overnight at 4 °C.

Techniques: Migration, In Vitro, Control, Flow Cytometry, Plasmid Preparation, shRNA, Quantitative RT-PCR, Cell Culture, Transfection, Expressing