Journal: Experimental and Therapeutic Medicine

Article Title: S100 calcium‑binding protein A16 suppresses the osteogenic differentiation of rat bone marrow mesenchymal stem cells by inhibiting SMAD family member 4 signaling

doi: 10.3892/etm.2024.12538

Figure Lengend Snippet: Inhibition of the MAPK/JNK pathway restores the osteogenic differentiation of BMSCs from OVX rats or S100A16-overexpressing wild-type rat BMSCs. (A) Western blot assay confirmed the effective downregulation of MAPK signaling in BMSCs isolated from OVX rats upon treatment with the MAPK inhibitor U0126. (B) Western blot assay confirmed the successful downregulation of JNK signaling in BMSCs isolated from OVX rats after treatment with the JNK inhibitor SP600125. (C) A comparison of Alizarin Red S staining results among the indicated groups. (D) Western blot assay confirmed the downregulation of MAPK signaling in S100A16-overexpressed BMSCs upon treatment with the MAPK inhibitor U0126. (E) Western blot assay confirmed the downregulation of JNK signaling in S100A16-overexpressed BMSCs after treatment with the JNK inhibitor SP600125. (F) Alizarin Red S staining of BMSCs transfected with either control empty plasmid or S100A16-overexpressing plasmid with or without treatment with MAPK inhibitor U0126 or JNK inhibitor SP600125. BMSCs, bone marrow mesenchymal stem cells; OVX, ovariectomized; p-, phosphorylated.

Article Snippet: For inhibitor treatments, both U0126 (cat. no. T21332; TargetMol Chemicals Inc.) and SP600125 (cat. no. T3109; TargetMol Chemicals Inc.) were used at a concentration of 10 μM.

Techniques: Inhibition, Western Blot, Isolation, Comparison, Staining, Transfection, Control, Plasmid Preparation, Immunohistochemistry, Over Expression

Journal: The Journal of Biological Chemistry

Article Title: KRAS G12V Enhances Proliferation and Initiates Myelomonocytic Differentiation in Human Stem/Progenitor Cells via Intrinsic and Extrinsic Pathways *

doi: 10.1074/jbc.M110.201848

Figure Lengend Snippet: Inhibition of ERK impairs proliferation while inhibition of p38 impairs proliferation and differentiation in CB CD34+ cells transduced with KRASG12V. A, Western blots of MiNR1 and KRASG12V-transduced CB CD34+ cells. B, UT7 cells were treated with PMA for 15 min, and cells were pretreated with the JNK inhibitor SP600125 as indicated after which lysates were prepared for Western blotting. C, Western blots on transduced cells as in A, but now cells were pretreated with the MEK inhibitor U0126 or the p38 inhibitor (SB203580) as indicated. D, proliferation of MiNR1 or KRASG12V cells in MS5 coculture in the presence of MEK inhibitor (U0126). Cultures were demi-depopulated weekly and cumulative cell counts are shown. In the right panels, the relative effects of the inhibitor are shown whereby the untreated MiNR1 and KRASG12V cell counts were normalized to 100% every week. E, As in D, but now MS5 cocultures were performed in the presence of p38 inhibitor (SB203580). F, FACS analysis of cultures described in E. G, FACS analysis of cultures described in E.

Article Snippet: The MEK inhibitor U0126 was from Promega Benelux B.V. Leiden, The Netherlands, the JNK inhibitor SP600125, and the p38 inhibitor SB203580 were obtained from VWR International (Roden, The Netherlands) and were used in concentrations as indicated in the text.

Techniques: Inhibition, Transduction, Western Blot

Journal: Cells

Article Title: Comparison of Differentiation Pattern and WNT/SHH Signaling in Pluripotent Stem Cells Cultured under Different Conditions

doi: 10.3390/cells10102743

Figure Lengend Snippet: Analysis of ESC differentiation in medium supplemented either with FCS or SR after modification of either canonical or noncanonical WNT pathway. ( A ) Expression of germ layer markers in control or IWR-1-treated EBs. ( B ) Expression of germ layer markers in control or SP600125-treated EBs. ( C ) Expression of germ layer markers in control or siRNA-transfected EBs. ( D ) Expression of germ layer markers in control or CHIR-treated EBs. β-actin was used as a reference gene. RQ = 1 for the expression level detected in a 13.5-day-old mouse embryo. The average ΔCq for reference sample is shown on each graph. Data presented as means of three independent experiments with standard deviations; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: To inhibit secretion of all WNTs or block either canonical or noncanonical WNT pathway EBs were cultured either in medium supplemented with 15% SR or 15% ESC-qualified FCS in the presence of either 5 μM IWP-2 (inhibitor of WNT secretion; Stem Cell Technologies, Vancouver, Canada), or 5 μM IWR-1 (canonical WNT pathway inhibitor; Stem Cell Technologies) or SP600125 (noncanonical WNT pathway inhibitor, JNK inhibitor; Stem Cell Technologies) for 2, 5 or 7 days.

Techniques: Modification, Expressing, Control, Transfection