Fig. 3

AS-IV inhibits senescent phenotypes in cultured astrocytes. Astrocytes were cultured for 10 days or 40 days in vitro (DIV) and then treated with AS-IV (1, 10, 50, 100, 200 μM) for 10 days. a, b Representative images of SA-β-gal activity (a) and percentage of SA-β-gal⁺ cells (b) (from three independent experiments). DAPI staining nucleus (blue). Scale bar 100 μm. c Cell viability was measured by CCK-8 assay (from five independent experiments). d, e Representative immunoblots (d) and quantitative analysis of p16^Ink4a (e) in astrocytes (from four independent experiments). f–k qPCR of p16^INK4a (f), CXCL1 (g), IL-1β (h), IL-6 (i), MMP3 (j), and MMP9 (k) mRNA levels (from four to six independent experiments). Quantified data are normalized to the control group (the control group value is equal to 1). The data shown are the mean ± SEM. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001. l Representative images of lamin B1 immunofluorescence (green) in GFAP⁺ astrocytes (red). m Quantification of relative lamin B1 fluorescence intensity in GFAP⁺ astrocytes. Quantified data are normalized to the control group (the control group value is equal to 100%). The data shown are the mean ± SEM from three independent experiments. ^*p < 0.05, ^***p < 0.001. DAPI staining nucleus (blue). Scale bar 25 μm. AS-IV, Astragaloside IV