p pyk2  (Cell Signaling Technology Inc)

Journal: Cells

Article Title: CD38 Inhibitor 78c Attenuates Pro-Inflammatory Cytokine Expression and Osteoclastogenesis in Macrophages

doi: 10.3390/cells13231971

Figure Lengend Snippet: Inhibition of CD38 by 78c inhibited podosome components (PI3K, Pyk2, Src, integrin β3, F-actin, paxillin, and talin) induced by RANKL. Murine bone marrow cells were treated with vehicle (diluted DMSO) or 78c (5 μM) with or without RANKL stimulation for four days. ( A ) Protein levels of p-PI3K, p-Pyk2, p-Src, CD38, and pan-actin protein levels were evaluated using Western blot. Protein densitometry of ( B ) p-PI3K, ( C ) p-Pyk2, ( D ) p-Src, and ( E ) CD38 were evaluated. ( F ) Protein levels of integrin β3, F-actin, paxillin, talin, and pan-actin protein levels were evaluated using Western blot. Protein densitometry of integrin β3 ( G ), F-actin ( H ), paxillin ( I ), and talin ( J ) were evaluated. Statistics were evaluated using an unpaired t -test with Welch’s correction ( n = 3, ns: no significance, * p < 0.05, ** p < 0.01).

Article Snippet: The antibodies to p-PI3K, p-ERK, p-JNK, p-p38, p-NF-κB p65, p-Src, p-Pyk2, integrin β3, and pan-actin were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Inhibition, Western Blot

Journal: Cells

Article Title: CD38 Inhibitor 78c Attenuates Pro-Inflammatory Cytokine Expression and Osteoclastogenesis in Macrophages

doi: 10.3390/cells13231971

Figure Lengend Snippet: Effect of a CD38 shRNA on osteoclastogenesis and bone resorption induced by RANKL. Murine bone marrow cells were treated with a control shRNA or a CD38 shRNA lentiviral vector (MOI 10). At 24 h after lentiviral infection, the cells were unstimulated or stimulated with RANLK for five days (in a 96-well cell culture plate for osteoclastogenesis assay), eight days (in a calcium phosphate-coated 48-well plate for bone resorption assay), or four days (for RT-PCR assay and Western blot assay). ( A ) Representative images show TRAP-stained cells at the 100× magnification view. Scale bars represent 50 μM. ( B ) Number of TRAP + multinucleated (more than three nuclei) osteoclasts/well (96-well) ( n = 4). ( C ) Total areas of osteoclast/image were quantified ( n = 4). ( D ) Representative images show osteoclast resorption pits at 200× magnification view. Scale bars represent 100 μM. ( E ) CD38 mRNA level, ( F ) Nfatc1 mRNA level, ( G ) Ctsk mRNA level, ( H ) Acp5 mRNA level, ( I ) Oscar mRNA level, ( J ) Ocstamp mRNA level, and ( K ) Dcstamp mRNA levels were quantified using RT-PCR and normalized by β-actin expression. Statistics were analyzed using an ordinary one-way ANOVA with Tukey’s multiple comparisons test ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001). ( L ) Protein levels of p-PI3K, p-Pyk2, p-Src, F-actin, integrin β3, paxillin, talin, CD38, and pan-actin protein levels were evaluated using Western blot.

Article Snippet: The antibodies to p-PI3K, p-ERK, p-JNK, p-p38, p-NF-κB p65, p-Src, p-Pyk2, integrin β3, and pan-actin were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: shRNA, Control, Plasmid Preparation, Infection, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Western Blot, Staining, Expressing

Journal: Cancer Medicine

Article Title: MCM8 promotes gastric cancer progression through RPS15A and predicts poor prognosis

doi: 10.1002/cam4.7424

Figure Lengend Snippet: MCM8/RPS15A axis promotes P38α, LYN, and p70S6K phosphorylation in GC. (A, B) Human Phospho‐Kinase Array Kit (ARY003C) was used to detect changes in phosphorylation of key proteins after RPS15A knockdown. (C, D) WB was applied to detect RPS15A, P53, LYN, P38α, c‐Jun, STAT1, STAT3, PYK2, p70S6K and their phosphorylated forms. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: MCM8 (1:500, Proteintech, Chicago, USA), RPS15A (1:2000, Proteintech, Chicago, USA), CDK4 (1:1000, Proteintech, Chicago, USA), MAPK14 (1:2000, Proteintech, Chicago, USA), CCND1 (1:750, CST, Boston, USA), SMAD3 (1:1000, Abcam, Cambridge, UK), SMAD4 (1:500, Santa Cruz, California, USA), ARAF (1:1000, CST, Boston, USA), RPS6KA1 (1:500, Proteintech, Chicago, USA), GAPDH (1:30000, Proteintech, Chicago, USA), P53 (1:3000, Proteintech, Chicago, USA), p‐P53 (1:2000, Proteintech, Chicago, USA), STAT3 (1:1500, CST, Boston, USA), p‐STAT3 (1:500, CST, Boston, USA), c‐Jun (1:1000, Proteintech, Chicago, USA), p‐c‐Jun (1:2000, CST, Boston, USA), p70S6K (1:1000, Affinity, Cincinnati, USA), p‐p70S6K (1:1000, Affinity, Cincinnati, USA), P38α(1:2000, Abcam, Cambridge, UK), p‐P38α(1:1000, Abcam, Cambridge, UK), LYN (1:1000, CST, Boston, USA), p‐LYN (1:1000, CST, Boston, USA), PYK2 (1:2000, Abcam, Cambridge, UK), p‐PYK2 (1:1000, CST, Boston, USA), STAT1 (1:1000, Proteintech, Chicago, USA), p‐STAT1 (1:2000, Abcam, Cambridge, UK).

Techniques: Knockdown