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. Author manuscript; available in PMC: 2022 Aug 9.
Published in final edited form as: Nat Cell Biol. 2020 Jan 6;22(1):74–86. doi: 10.1038/s41556-019-0441-z

Extended Data Fig. 9. SR9009 affects collagen fibril assembly.

Extended Data Fig. 9

a, Immunofluorescence analysis showed more collagen-I fibres assembled by the fibroblasts of ClockΔl9 mice. N = 3 biological repeats. Bars 10 μm. b, Kill curve of SR9009 showing that concentration of 10 μM (indicated with arrow) is well tolerated by WT and Δ19 fibroblasts. c,d, Luminometry recordings of circadian rhythms of dexamethasone-synchronized fibroblasts isolated from PER2::Luciferase (WT) and ClockΔ19::PER2::Luciferase (Δ19) mice. SR2.5, 2.5 μM SR9009. SR10, 10 μM SR9009. Arrow indicates addition of SR9009. e,f, Immunofluorescence analysis of wild-type and ClockΔ19 fibroblasts either treated with 10 μM SR9009 or mock-treated with dimethylsulfoxide (Ctrl). Blue, DAPI. Red, anti-collagen-I antibody. Number of DAPI-stained nuclei (that is number of cells) and number of fibres were blind scored from randomly- selected regions from n = 3 biological repeats by 3 observers. Number of collagen fibres per cell is shown for control and SR9009-treated fibroblasts. Fold change from qPCR analysis of gene expression is shown for control and SR9009-treated fibroblasts. *p = 0.0127, WT Ctrl vs WT + SR9009; ***p = 0.0005 Δ19Ctrl vs Δ19SR9009. Q-PCR analysis for SR9009-treated WT cells: *p = 0.0476, Sec61a2; *p = 0.0072, Mia3; **p = 0.0042, Serpinh1; *p = 0.0226, Pde4d. Q-PCR analysis for SR9009-treated Δ19 cells: **p = 0.0072, Vps33b; two-tailed paired t-test, n = 4 biological replicates. Bars show s.e.m.