Journal: Stem Cells International

Article Title: Palovarotene Can Attenuate Heterotopic Ossification Induced by Tendon Stem Cells by Downregulating the Synergistic Effects of Smad and NF- κ B Signaling Pathway following Stimulation of the Inflammatory Microenvironment

doi: 10.1155/2022/1560943

Figure Lengend Snippet: The primer sequences used in this study.

Article Snippet: The proteins in the film were then transferred to a polyvinylidene difluoride (PVDF) membrane, using 5% skimmed milk powder at room temperature for 1 h. OCN (Servicebio, #GB11233) or RUNX2 (Bioss, #bs1134R) or SOX9 (Bioss, #bs4177R), beta actin (Servicebio, #GB11001) or P65 (Abcam, #ab179463) or Smad1/5 (Abcam, #ab38449) or Id1 (Proteintech, #18475-1--AP) or p-Smad1/5 (Abcam, #ab51451) were added to the blocking solution to dilute to the desired concentration and were incubated overnight with the membrane at 4°C.

Techniques:

Journal: Stem Cells International

Article Title: Palovarotene Can Attenuate Heterotopic Ossification Induced by Tendon Stem Cells by Downregulating the Synergistic Effects of Smad and NF- κ B Signaling Pathway following Stimulation of the Inflammatory Microenvironment

doi: 10.1155/2022/1560943

Figure Lengend Snippet: The inflammatory microenvironment promotes osteogenic differentiation of TSCs, and palovarotene can influence osteogenesis. (a) CCK8 proliferation assay was used to determine the dose range of palovarotene. Palovarotene at 0.5, 1, and 2 μ M concentration had less effect on TSCs proliferation. Coculture system of activated macrophages and TSCs was constructed. (b) ALP and ARS staining in TSCs after coculture with macrophages. Data are means ± SD ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01. # p < 0.05 and ## p < 0.01. Scale bar = 50 μ m. (c) Western blot analysis and relevant quantitative analysis of OCN, SOX9, and RUNX2. Data are means ± SD ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01. # p < 0.05 and ## p < 0.01. (d) The mRNA expression of OCN, SOX9, RUNX2, MMP-9, IL-1 β , IFN- γ , and TNF- α . Data are means ± SD ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01. # p < 0.05 and ## p < 0.01.

Article Snippet: The proteins in the film were then transferred to a polyvinylidene difluoride (PVDF) membrane, using 5% skimmed milk powder at room temperature for 1 h. OCN (Servicebio, #GB11233) or RUNX2 (Bioss, #bs1134R) or SOX9 (Bioss, #bs4177R), beta actin (Servicebio, #GB11001) or P65 (Abcam, #ab179463) or Smad1/5 (Abcam, #ab38449) or Id1 (Proteintech, #18475-1--AP) or p-Smad1/5 (Abcam, #ab51451) were added to the blocking solution to dilute to the desired concentration and were incubated overnight with the membrane at 4°C.

Techniques: Proliferation Assay, Concentration Assay, Construct, Staining, Western Blot, Expressing

Journal: Stem Cells International

Article Title: Palovarotene Can Attenuate Heterotopic Ossification Induced by Tendon Stem Cells by Downregulating the Synergistic Effects of Smad and NF- κ B Signaling Pathway following Stimulation of the Inflammatory Microenvironment

doi: 10.1155/2022/1560943

Figure Lengend Snippet: Smad and NF- κ B signaling pathways play a synergistic role in HO formation, and palovarotene inhibits HO formation by blocking the Smad and NF- κ B signaling pathways. (a) The mRNA expression of OCN, SOX9, RUNX2, Smad1, Smad5, and ID1. Data are means ± SD ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01. # p < 0.05 and ## p < 0.01. (b) Immunofluorescence against ID1 ( n = 3). Scale bar = 25 μ m. (c) Western blot analysis and relevant quantitative analysis of p-p65 and ID1. Data are means ± SD ( n = 3) ∗ p < 0.05 and ∗∗ p < 0.01. # p < 0.05 and ## p < 0.01. (d) Immunofluorescence against p65 ( n = 3). Scale bar = 25 μ m. (e) Western blot analysis and relevant quantitative analysis of p65 and immunofluorescence against P65 in the previous coculture model series experiments ( n = 3). Data are means ± SD ( n = 3). ∗ p < 0.05 and ∗∗ p < 0.01. # p < 0.05 and ## p < 0.01. Scale bar = 25 μ m.

Article Snippet: The proteins in the film were then transferred to a polyvinylidene difluoride (PVDF) membrane, using 5% skimmed milk powder at room temperature for 1 h. OCN (Servicebio, #GB11233) or RUNX2 (Bioss, #bs1134R) or SOX9 (Bioss, #bs4177R), beta actin (Servicebio, #GB11001) or P65 (Abcam, #ab179463) or Smad1/5 (Abcam, #ab38449) or Id1 (Proteintech, #18475-1--AP) or p-Smad1/5 (Abcam, #ab51451) were added to the blocking solution to dilute to the desired concentration and were incubated overnight with the membrane at 4°C.

Techniques: Blocking Assay, Expressing, Immunofluorescence, Western Blot

Journal: Journal of translational medicine

Article Title: RUNX1 knockdown induced apoptosis and impaired EMT in high-grade serous ovarian cancer cells.

doi: 10.1186/s12967-023-04762-8

Figure Lengend Snippet: Fig. 5 RUNX1 knockdown affects EMT in ovarian cancer cells via the EGFR-AKT-STAT3 axis. Western blot detected the signaling molecules change pattern in the RUNX1 KD cells that involved in the EMT related signaling pathway (A and B). Western blot detected the signaling molecules change pattern after Ro5-3335 (a RUNX1 inhibitor) treatment that involved in the EMT related signaling pathway (C and D). Western blot detected of EMT-related molecules in RUNX1 knockdown cell lines (E and F). Western blot detected of EMT-related molecules after Ro5-3335 treatment (G and H). The expression of EMT-related molecules was analyzed by immunofluorescence in RUNX1 knockdown cell lines (I and J). *p < 0.05, **p < 0.01 and ***p < 0.001 as compared with control cells expressing a scramble shRNA control, paired t test

Article Snippet: Sections were incubated overnight with RUNX1 antibody (Cusabio, China) at 4 °C.

Techniques: Knockdown, Western Blot, Expressing, Immunofluorescence, Control, shRNA

Journal: Bioscience Reports

Article Title: Down-regulation of lncRNA-NEF indicates poor prognosis in intrahepatic cholangiocarcinoma

doi: 10.1042/BSR20181573

Figure Lengend Snippet: LncRNA-NEF and RUNX1 mRNA were differentially expressed in tumor tissues and adjacent healthy tissues of IHCC patients Compared with adjacent healthy tissues, expression levels of lncRNA-NEF were significantly increased ( A ), while expression levels of RUNX1 mRNA were significantly up-regulated ( B ) in tumor tissues of IHCC patients. * P <0.05.

Article Snippet: Primary antibodies included rabbit anti-human RUNX1 (ab35962, 1:1400, Abcam) and GAPDH (ab9485, 1:1400, Abcam).

Techniques: Expressing

Journal: Bioscience Reports

Article Title: Down-regulation of lncRNA-NEF indicates poor prognosis in intrahepatic cholangiocarcinoma

doi: 10.1042/BSR20181573

Figure Lengend Snippet: Expression levels of lncRNA-NEF and RUNX1 were significantly and reversely correlated in tumor tissues but not in adjacent healthy tissues. Pearson Correlation Coefficient revealed a significantly reverse correlation between the expression levels of lncRNA-NEF and RUNX1 mRNA in tumor tissues ( A ) but not in healthy tissues ( B ).

Article Snippet: Primary antibodies included rabbit anti-human RUNX1 (ab35962, 1:1400, Abcam) and GAPDH (ab9485, 1:1400, Abcam).

Techniques: Expressing

Journal: Bioscience Reports

Article Title: Down-regulation of lncRNA-NEF indicates poor prognosis in intrahepatic cholangiocarcinoma

doi: 10.1042/BSR20181573

Figure Lengend Snippet: LncRNA-NEF overexpression led to inhibited RUNX1 expression in IHCC cells LncRNA-NEF overexpression led to significantly inhibited expression of RUNX1 in cells of both HuCCT1 and TFK-1 human IHCC cell lines ( A ). In contrast, RUNX1 overexpression did not significantly alter the expression of lncRNA-NEF in those cells ( B ) (* P <0.05).

Article Snippet: Primary antibodies included rabbit anti-human RUNX1 (ab35962, 1:1400, Abcam) and GAPDH (ab9485, 1:1400, Abcam).

Techniques: Over Expression, Expressing

Journal: Bioscience Reports

Article Title: Down-regulation of lncRNA-NEF indicates poor prognosis in intrahepatic cholangiocarcinoma

doi: 10.1042/BSR20181573

Figure Lengend Snippet: LncRNA-NEF overexpression inhibited IHCC cell migration and invasion possibly through RUNX1 LncRNA-NEF overexpression led to significantly inhibited, while RUNX1 led to significantly promoted migration ( A ) and invasion ( B ) of cells of both HuCCT1 and TFK-1 human IHCC cell lines. In addition, RUNX1 overexpression partially compensate the inhibitory effects of lncRNA-NEF overexpression on cancer cell migration and invasion (* P <0.05).

Article Snippet: Primary antibodies included rabbit anti-human RUNX1 (ab35962, 1:1400, Abcam) and GAPDH (ab9485, 1:1400, Abcam).

Techniques: Over Expression, Migration