Extended Data Fig. 5: Effects of BMP4 in cocktail-induced 3D spheroids.
From: BMP4 and Gremlin 1 regulate hepatic cell senescence during clinical progression of NAFLD/NASH
(a, b) Representative immunofluorescence images of 3D spheroids (IHH/LX-2, 24:1) treated with cocktail (oleic acid (200 µM) + TGFβ1 (5 ng/ml) + TNFα (0.2 nM)) in presence or absence of BMP4/GREM1 for 48 h, stained for ORO (red) (a), COL1A1 (green) (b), and nuclei (DAPI, blue). Scale bar represents 50 µm. (c, d) Bar graph displays fluorescence intensities of ORO (c) and COL1A1 (d) quantified using ImageJ and normalized to number of nuclei (n = 20-30 spheroids from 3 different experiments). Values are means ± SEM. Statistical significance was determined using Kruskal-Wallis with post-hoc Dunn’s test. a.u., arbitrary unit.
