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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: Functional crosstalk between mTORC1/p70S6K pathway and heterochromatin organization in stress-induced senescence of MSCs

Fig. 5

mTORC1 pathway cross talks with heterochromatin organization during stress-induced senescence. a 1 × 104 hUC-MSCs (p8-p9) were seeded in coverslips and treated with Dox with or without rapamycin. Cells were collected after 24 h. Representative images and quantification of immunofluorescence staining of HP1γ and H3K9me3 in control or rapamycin-treated hUC-MSCs in response to Dox (scale bar = 10 μm). Quantification is shown on the right panel, mean ± SEM of values from three independent experiments with triplicate wells analyzed on 6–8 cells/field from five different fields.*p < 0.05; **p < 0.01 by one-way ANOVA with Tukey’s post hoc test. b 1 × 104 hUC-MSCs (p5-p7) were seeded coverslips and transfected with siRNAs. Twenty-four hours later, the cells were treated with Dox and collected 1 day afterward. Immunofluorescent staining of heterochromatin marks H3K9me3 and HP1γ shows that knockdown of p70S6K increases the H3K9me3 foci and HP1γ intensity. Quantification is shown on the right panel, mean ± SEM of values from three independent experiments with triplicate wells analyzed on 6–8 cells/field from five different fields.*p < 0.05; **p < 0.01, ***p < 0.001 by one-way ANOVA with Tukey’s post hoc test. c Representative Western blot shows that knockdown of p70S6K increases the expression levels of H3K9me3 and HP1γ. Data are presented as the mean ± SEM. **p < 0.01; ***p < 0.001 by one-way ANOVA with Tukey’s post hoc test. d 1 × 104 hUC-MSCs (p8-p9) were seeded in coverslips and treated with Dox with or without MHY1485. Cells were collected after 24 h. Representative images and quantification of immunofluorescence staining of HP1γ and H3K9me3 in control or MHY1485-treated hUC-MSCs in response to Dox (scale bar = 10 μm). Quantification is shown on the right panel, mean ± SEM of values from three independent experiments with triplicate wells analyzed on 6–8 cells/field from five different fields.*p < 0.05; **p < 0.01;***p < 0.001 by one-way ANOVA with Tukey’s post hoc test. e Representative Western blot shows that MHY1485 reduces the expression levels of H3K9me3 and HP1γ. Quantification data is shown below. Data are achieved from three independent experiments. *p < 0.05; **p < 0.01;***p < 0.001 by one-way ANOVA with Tukey’s post hoc test

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