Extended Data Fig. 4. Inhibition of IL11 signalling reduces replicative senescence, inflammation, ageing biomarkers, and metabolic decline in human cardiac fibroblasts.
a-c Data for HCF at passage 4 (P4), 7, 10, and 14 that had been passaged in the presence of either IgG or anti-IL11RA (X209; 2 µg/ml) from P2. a WB of total and p-ERK1/2, p-p90RSK, p-LKB1, p-AMPK, p-mTOR, p-p70S6K, p-S6RP, p-NFκB, p-STAT3, p16, p21, PCNA, Cyclin D, and GAPDH (n = 6/group). b Immunofluorescence images (scale bars, 100 μm; representative datasets from n = 7/group) and quantification of intensity/area (n = 14/group) for p16 and p21 staining. c IL11, IL6 and IL8 levels in the supernatant based on ELISA (n = 6/group). d WB showing the expression levels of p16, p21, and GAPDH from HCFs P4 that were stimulated for 8, 24, 48, and 72 h with media collected from HCFs P14 that had been grown and passaged in the presence of either IgG or anti-IL11RA (X209; 2 µg/ml) from P2 (n = 4/group). e Telomere length (n = 6/group) and f mtDNA copy number (n = 6/group) and seahorse assay (n = 8/group) showing g mitochondrial oxygen consumption rate (OCR), h changes in OCR during basal respiration and ATP production states, and i oxidative and glycolytic energy phenotypes at baseline in HCFs P4 and P14 either untreated or in the presence of either IgG or anti-IL11RA (X209; 2 µg/ml). b, c, e-i Data are shown as mean ± SD. b, c Two-way ANOVA with Sidak’s correction, e, f, h one-way ANOVA with Tukey’s correction. For gel source data, see Supplementary Fig. 1.