Journal: Experimental and Therapeutic Medicine

Article Title: Ajuga decumbens stimulates mesenchymal stem cell differentiation and regenerates cartilage in a rabbit osteoarthritis model

doi: 10.3892/etm.2018.5981

Figure Lengend Snippet: Chondrocyte differentiation from hMSCs. hMSCs were cultured for 3 weeks with GM or CD and different concentrations of EADE. Cells were subjected to Alcian blue staining and the absorbance was measured at 630 nm. *P<0.05 and **P<0.01. hMSCs, human mesenchymal stem cells; GM, normal growth medium; CD, chondrogenic differentiation medium; EADE, extra Ajuga decumbens extract.

Article Snippet: Human chondrocytes (HC) derived from normal human femoral cartilage were obtained from Cell Applications, Inc. (Merck KGaA).

Techniques: Cell Culture, Staining

Journal: Experimental and Therapeutic Medicine

Article Title: Ajuga decumbens stimulates mesenchymal stem cell differentiation and regenerates cartilage in a rabbit osteoarthritis model

doi: 10.3892/etm.2018.5981

Figure Lengend Snippet: Effect of EADE on PGE2 production in chondrocytes. Human chondrocytes were stimulated with IL-1β (1 ng/ml) and treated with 10 or 100 µg/ml EADE. The concentration of PGE2 was significantly increased by IL-1β stimulation, and EADE significantly suppressed this increase. *P<0.05. IL, interleukin; EADE, extra Ajuga decumbens extract; PGE2, prostaglandin E2.

Article Snippet: Human chondrocytes (HC) derived from normal human femoral cartilage were obtained from Cell Applications, Inc. (Merck KGaA).

Techniques: Concentration Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Glycogen synthase kinase-3beta facilitates IFN-gamma-induced STAT1 activation by regulating Src homology-2 domain-containing phosphatase 2.

doi: 10.4049/jimmunol.0804033

Figure Lengend Snippet: FIGURE 2. IFN- regulates Akt and Pyk2, which then activates GSK-3 and inflammation. A and C, RAW264.7 cells were treated with IFN- for the indicated time periods. We used Western blotting to determine phosphoryla- tion of GSK-3 (Ser9 and Tyr216) and Akt (Ser473). B, D, and F, With or without BIO, OA, or Sph-24 pretreatment for 0.5 h, RAW264.7 cells were then treated with IFN-. Western blotting was used to determine phosphory- lation of GS (Ser641) in cells 2 h posttreatment, and Akt (Ser473) and GSK-3 (Ser9) in cells 0.5 h posttreatment. E, RAW264.7 cells were treated with IFN- for the indicated time periods. We used a kit to determine SMase activity. Percentages relative to those of the control are shown. G, Western blotting was used to determine the time kinetics of phosphorylation of Pyk2 (Tyr402) in IFN--stimulated RAW264.7 cells. H–J, With or without A9 pretreatment for 0.5 h, RAW264.7 cells were then treated with IFN-. Western blotting was used to determine phosphorylation of GSK-3 (Ser9) 0.25 h posttreatment and GSK-3 (Tyr216) 2 h posttreatment. Griess reagent was used to determine NO production 24 h posttreatment. TNF- production was determined using ELISA 6 h posttreatment. For Western blot analysis, -actin was the internal control. Data shown are representative of three individual experiments. For the activity assay, nitrite detection, and ELISA, the data are means SD obtained from three individual cultures. , p 0.05 compared with the untreated or the IFN- group.

Article Snippet: Abs against phospho-GSK-3 (Ser9), phospho-GSK-3 (Tyr216), GSK-3 / , phospho-GS (Ser641), GS, phospho-Akt (Ser473), Akt, phospho-Pyk2 (Tyr402), Pyk2, phospho-STAT1 (Tyr701), phospho-STAT1 (Ser727), STAT1, phospho-Jak2 (Tyr1007/1008), Jak2, phospho-Src (Tyr416), Src, phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-p38 MAPK (Thr180/Tyr182), p38 MAPK, phospho-SHP2 (Tyr542), SHP2, SOCS1, and SOCS3 were purchased from Cell Signaling Technology.

Techniques: Western Blot, Activity Assay, Control, Phospho-proteomics, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Glycogen synthase kinase-3beta facilitates IFN-gamma-induced STAT1 activation by regulating Src homology-2 domain-containing phosphatase 2.

doi: 10.4049/jimmunol.0804033

Figure Lengend Snippet: FIGURE 3. IFN- induces PC-PLC-, PKC-, and Src-regulated GSK-3 activation and inflammation. RAW264.7 cells were pretreated with D609, Go¨6976, or PP1 for 0.5 h and then treated with IFN-. We used Western blotting to determine the phosphorylation of Pyk2 (Tyr402) and GSK-3 (Tyr216) 2 h posttreatment and GSK-3 (Ser9) 0.5 h posttreatment and iNOS expression 24 h posttreatment (A–C). Griess reagent and ELISA were used to determine NO (D) 24 h posttreatment and TNF- (E) pro- duction 6 h posttreatment. For Western blot analysis, -actin was the in- ternal control. Data shown are representative of three individual experi- ments. The OD of Pyk2 (Tyr402)/total Pyk2 has been clarified using ImageJ software. For nitrite detection and ELISA, the data are means SD ob- tained from three individual cultures. , p 0.05 compared with the IFN- group.

Article Snippet: Abs against phospho-GSK-3 (Ser9), phospho-GSK-3 (Tyr216), GSK-3 / , phospho-GS (Ser641), GS, phospho-Akt (Ser473), Akt, phospho-Pyk2 (Tyr402), Pyk2, phospho-STAT1 (Tyr701), phospho-STAT1 (Ser727), STAT1, phospho-Jak2 (Tyr1007/1008), Jak2, phospho-Src (Tyr416), Src, phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-p38 MAPK (Thr180/Tyr182), p38 MAPK, phospho-SHP2 (Tyr542), SHP2, SOCS1, and SOCS3 were purchased from Cell Signaling Technology.

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Control, Software

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Glycogen synthase kinase-3beta facilitates IFN-gamma-induced STAT1 activation by regulating Src homology-2 domain-containing phosphatase 2.

doi: 10.4049/jimmunol.0804033

Figure Lengend Snippet: FIGURE 4. IFN- induces Jak2-regulated activation of PC-PLC, Pyk2, and GSK-3 independently of IFNGR1. A, Splenocytes harvested from IFNGR1 wild-type (IFNGR1 WT) and IFNGR1-deficient (IFNGR1/) C57BL/6 mice were treated with IFN- for 0.5 h. We used a kit to deter- mine the activity of PC-PLC. Percentages relative to those of the control are shown. B, Western blotting was used to determine phosphorylation of STAT1 (Tyr701) 1 h posttreatment and Pyk2 (Tyr402) and GSK-3 (Tyr216) 3 h posttreatment. C, RAW264.7 cells were pretreated with AG490 for 0.5 h and then treated with IFN- for 0.5 h. The activity of PC-PLC was determined using a kit. Percentages relative to those of the control are shown. D, Western blotting was used to determine phosphorylation of STAT1 (Tyr701) and Jak2 (Tyr1007/1008) 1 h posttreatment and Pyk2 (Tyr402) and GSK-3 (Tyr216) 3 h posttreatment. For the activity assay, the data are means SD obtained from three individual cultures. , p 0.05 compared with the untreated or the IFN- group. For Western blot analysis, -actin was the internal control. Data shown are representative of three individual experiments.

Article Snippet: Abs against phospho-GSK-3 (Ser9), phospho-GSK-3 (Tyr216), GSK-3 / , phospho-GS (Ser641), GS, phospho-Akt (Ser473), Akt, phospho-Pyk2 (Tyr402), Pyk2, phospho-STAT1 (Tyr701), phospho-STAT1 (Ser727), STAT1, phospho-Jak2 (Tyr1007/1008), Jak2, phospho-Src (Tyr416), Src, phospho-ERK1/2 (Thr202/Tyr204), ERK1/2, phospho-p38 MAPK (Thr180/Tyr182), p38 MAPK, phospho-SHP2 (Tyr542), SHP2, SOCS1, and SOCS3 were purchased from Cell Signaling Technology.

Techniques: Activation Assay, Activity Assay, Control, Western Blot, Phospho-proteomics