Fig. 6: HPP-9 is a long-acting Hedgehog pathway inhibitor. | Nature Communications

Fig. 6: HPP-9 is a long-acting Hedgehog pathway inhibitor.

From: Targeted protein degradation reveals BET bromodomains as the cellular target of Hedgehog pathway inhibitor-1

Fig. 6

a SHH-GFP cells were treated for 25 h with DMSO, 1 μM HPI-1, or 1 μM HPP-9, before the medium was changed to various dilutions of ShhN-conditioned medium for 27 h. Nuclear GFP levels were quantified using fluorescence microscopy. Representative curves of three independent experiments are shown, with 9–16 images analyzed per condition. b GLI1 and GLI2 levels of cells pre-incubated with DMSO or 1 μM HPP-9 were determined. Representative blots of three independent experiments are shown. c, d The effect of 1 μM HPP-9 or HPI-1 (pre-)incubation on GLI2 ciliary trafficking was assessed through fluorescence microscopy. Representative images for GLI2 trafficking are shown in (c), and all data is quantified in (d). N independent experiments as indicated in the bars, n = 300–500 cilia analyzed per condition. Mean ± SD is plotted, two-way ANOVA, p as indicated. Scalebar 2 μm. e SHH-GFP cells were incubated with the indicated compounds for 25 h, before the medium was changed to ShhN-conditioned medium with or without competitors for 27 h. Nuclear BRD protein levels were determined using high-content fluorescence microscopy. Data shown is from N independent experiments as indicated, with n = 9–16 images analyzed per condition in each experiment. Source data are provided as a Source Data file.

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