Mouse GLI-2 Antibody

Detection of Mouse GLI-2 by Western Blot

HPP-9 is a long-acting Hedgehog pathway inhibitor.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. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse GLI-2 by Western Blot

Biological evaluation of HPP-9 and its inactive analog.a Chemical structures of HPP-9 and the methylated analog inact-HPP-9. b Representative micrographs showing the dose-dependent inhibition of an Shh-driven GFP reporter by HPP-9, resulting in the dose-response curves shown in (c). Scalebar 30 μm. b, c Curves and images are representative of N independent experiments (as indicated in the table below the graph), with n = 9 or 18 images analyzed per experiment. d, e NIH-3T3 cells were incubated with increasing concentrations of HPP-9 or HPI-1 in the presence of ShhN and lysates were probed for GLI1, GLI2, and GLI3. d shows a representative immunoblot illustrating that both compounds inhibit GLI1 and GLI2 but have no effect on GLI3 processing. e Quantification (mean ± SEM) of GLI1 (N = 3) and GLI2 full-length levels (N = 2) of N independent experiments. f–h qPCR for Ptch1 (f), Gli1 (g), and Gli2 (h) shows that HPP-9 reduces the expression of Hh pathway target genes, while also decreasing basal Gli2 transcript levels without affecting the fold-induction upon pathway stimulation (−: no ShhN, +: with ShhN). Data shown is the mean (f) or mean ± SD (g, h) for N independent experiments as indicated in the bar. f, g One-way ANOVA with Dunnett’s test, p as indicated in the graph, compared to DMSO + ShhN. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse GLI-2 by Western Blot

Biological evaluation of HPP-9 and its inactive analog.a Chemical structures of HPP-9 and the methylated analog inact-HPP-9. b Representative micrographs showing the dose-dependent inhibition of an Shh-driven GFP reporter by HPP-9, resulting in the dose-response curves shown in (c). Scalebar 30 μm. b, c Curves and images are representative of N independent experiments (as indicated in the table below the graph), with n = 9 or 18 images analyzed per experiment. d, e NIH-3T3 cells were incubated with increasing concentrations of HPP-9 or HPI-1 in the presence of ShhN and lysates were probed for GLI1, GLI2, and GLI3. d shows a representative immunoblot illustrating that both compounds inhibit GLI1 and GLI2 but have no effect on GLI3 processing. e Quantification (mean ± SEM) of GLI1 (N = 3) and GLI2 full-length levels (N = 2) of N independent experiments. f–h qPCR for Ptch1 (f), Gli1 (g), and Gli2 (h) shows that HPP-9 reduces the expression of Hh pathway target genes, while also decreasing basal Gli2 transcript levels without affecting the fold-induction upon pathway stimulation (−: no ShhN, +: with ShhN). Data shown is the mean (f) or mean ± SD (g, h) for N independent experiments as indicated in the bar. f, g One-way ANOVA with Dunnett’s test, p as indicated in the graph, compared to DMSO + ShhN. Source data are provided as a Source Data file. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Mouse GLI-2 by Western Blot

HPP-9 is a long-acting Hedgehog pathway inhibitor.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. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37393376), licensed under a CC-BY license. Not internally tested by R&D Systems.