Fig. 7
FUS silencing via ION363 treatment ex vivo in IPF PCLs affects multiple pathways. The genes depicted in the volcano plot in Fig. 6f are tabulated in (a) (downregulated) and (b) (upregulated), with the respective FDR and log2FC values in ION363-treated IPF PCLs. c IPF PCLs were treated with scramble ASO (Scr) or FUS-ASO (ION363) followed by LysoTracker Red staining on PCLs, and live imaging of the treated PCLs was performed at the indicated times. The brightfield images depicted here are in combination with the red fluorescence signal of LysoTracker Red. Pictures and analysis of n = 1 IPF PCLs are shown. d Representative images showing immunostaining for COL1A1 (red) in IPF PCLs treated with Scr or ION363. DAPI was used to stain the nuclei. Scale bar = 300 µm. The graph on the right side represents the COL1A1 mean fluorescence intensity (MFI) in the indicated groups. e, f HD PCLs were treated with a profibrotic cocktail (PFC) or PFC in combination with ION363, as shown here by the analysis of FUS mRNA via qRT‒PCR. e Immunofluorescence staining for FUS (red) or α-SMA (green). Nuclei were stained with DAPI. Scale bar for: 10× images = 20 µm, 40× images = 50 µm. f. The quantification of the FUS MFI is shown, with pictures and analysis from n = 2 HD PCLS. P-value: **P < 0.01
