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rabbit anti usp39 antibody (Proteintech)

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Proteintech rabbit anti usp39 antibody

Fig. 1 <t>USP39</t> is localized in nucleolus and undergoes LLPS. A, Representative images of immunofluorescence assay showing cellular localization of GFP-USP39 fusion protein expressed in HeLa cells with or without treatment of 3% 1, 6-hexanediol (1,6-HD) for 7 min. B, Analysis of intrinsic disorder tendency of USP39 using PhaSepDB database. Below schematic represents corresponding domains of USP39. C, FRAP analysis in HeLa cells transfected with GFP-USP39 showing the recovery of the condensate after photobleaching. D, Schematic illustration of sub-nucleolar compartments, including the fibrillar center (FC), dense fibrillar component (DFC), and granular component (GC), with indicated markers being RPA40, FBL and NPM1 respectively. E, Immunofluorescence analysis of colocalization of GFP-tagged USP39 with NPM1, RAP40, and FBL in HeLa cells. Diagram curves on the right show quan tification. Scale bar, 10 μm

Rabbit Anti Usp39 Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+anti+usp39/USP39+Antibody/pm39885503-58-6-9
Average 93 stars, based on 30 article reviews

rabbit anti usp39 antibody - by Bioz Stars, 2026-09

93/100 stars

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1) Product Images from "USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression."

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

Journal: Cell communication and signaling : CCS

doi: 10.1186/s12964-025-02059-5

Fig. 1 USP39 is localized in nucleolus and undergoes LLPS. A, Representative images of immunofluorescence assay showing cellular localization of GFP-USP39 fusion protein expressed in HeLa cells with or without treatment of 3% 1, 6-hexanediol (1,6-HD) for 7 min. B, Analysis of intrinsic disorder tendency of USP39 using PhaSepDB database. Below schematic represents corresponding domains of USP39. C, FRAP analysis in HeLa cells transfected with GFP-USP39 showing the recovery of the condensate after photobleaching. D, Schematic illustration of sub-nucleolar compartments, including the fibrillar center (FC), dense fibrillar component (DFC), and granular component (GC), with indicated markers being RPA40, FBL and NPM1 respectively. E, Immunofluorescence analysis of colocalization of GFP-tagged USP39 with NPM1, RAP40, and FBL in HeLa cells. Diagram curves on the right show quan tification. Scale bar, 10 μm

Figure Legend Snippet: Fig. 1 USP39 is localized in nucleolus and undergoes LLPS. A, Representative images of immunofluorescence assay showing cellular localization of GFP-USP39 fusion protein expressed in HeLa cells with or without treatment of 3% 1, 6-hexanediol (1,6-HD) for 7 min. B, Analysis of intrinsic disorder tendency of USP39 using PhaSepDB database. Below schematic represents corresponding domains of USP39. C, FRAP analysis in HeLa cells transfected with GFP-USP39 showing the recovery of the condensate after photobleaching. D, Schematic illustration of sub-nucleolar compartments, including the fibrillar center (FC), dense fibrillar component (DFC), and granular component (GC), with indicated markers being RPA40, FBL and NPM1 respectively. E, Immunofluorescence analysis of colocalization of GFP-tagged USP39 with NPM1, RAP40, and FBL in HeLa cells. Diagram curves on the right show quan tification. Scale bar, 10 μm

Techniques Used: Immunofluorescence, Transfection

Fig. 2 USP39 IDR domain is required for LLPS. A, Schematic representation of USP39 and its truncated mutants containing or deleting IDR domain. B, Representative images of immunofluorescence assay showing the localization of GFP-tagged USP39 with full length and the 5 truncated versions of GFP- USP39 in HeLa cells. C, FRAP assay in HeLa cells transfected with GFP-USP39-N(1-103) showing the recovery of bleached droplet foci. D, Representative images of RC and fluorescence microscopy showing in vitro droplet formation of purified GFP vector and GFP-USP39-N(1-103) fusion protein both at 60 µM in the presence of PEG 4000 and 150 mM of NaCl. E, Representative images of RC and fluorescence microscopy showing in vitro droplet formation of purified GFP-USP39-N(1-103) fusion protein (60 µM) with escalating molecular weights of the PEG crowders as indicated. Scale bar, 10 μm

Figure Legend Snippet: Fig. 2 USP39 IDR domain is required for LLPS. A, Schematic representation of USP39 and its truncated mutants containing or deleting IDR domain. B, Representative images of immunofluorescence assay showing the localization of GFP-tagged USP39 with full length and the 5 truncated versions of GFP- USP39 in HeLa cells. C, FRAP assay in HeLa cells transfected with GFP-USP39-N(1-103) showing the recovery of bleached droplet foci. D, Representative images of RC and fluorescence microscopy showing in vitro droplet formation of purified GFP vector and GFP-USP39-N(1-103) fusion protein both at 60 µM in the presence of PEG 4000 and 150 mM of NaCl. E, Representative images of RC and fluorescence microscopy showing in vitro droplet formation of purified GFP-USP39-N(1-103) fusion protein (60 µM) with escalating molecular weights of the PEG crowders as indicated. Scale bar, 10 μm

Techniques Used: Immunofluorescence, FRAP Assay, Transfection, Fluorescence, Microscopy, In Vitro, Purification, Plasmid Preparation

Fig. 3 The definite condition of USP39 droplet formation was detected in serial concentration gradients. A and B, Diagram showing in vitro droplet formation of purified GFP-USP39-N(1-103) in the absence (A) or presence (B) of PEG 4000 with NaCl concentrations at 150, 200, 250, and 300 mM. The concentration of fusion protein added in buffer were as indicated. C, Representative images exhibiting droplet formation of GFP-USP39-N(1-103) fusion protein at the concentrations of 3, 6, 12, 20, 30 and 60 µM with NaCl concentrations ranging from 150 to 300 mM as illustrated in b. Scale bar, 10 μm

Figure Legend Snippet: Fig. 3 The definite condition of USP39 droplet formation was detected in serial concentration gradients. A and B, Diagram showing in vitro droplet formation of purified GFP-USP39-N(1-103) in the absence (A) or presence (B) of PEG 4000 with NaCl concentrations at 150, 200, 250, and 300 mM. The concentration of fusion protein added in buffer were as indicated. C, Representative images exhibiting droplet formation of GFP-USP39-N(1-103) fusion protein at the concentrations of 3, 6, 12, 20, 30 and 60 µM with NaCl concentrations ranging from 150 to 300 mM as illustrated in b. Scale bar, 10 μm

Techniques Used: Concentration Assay, In Vitro, Purification

Fig. 4 Restoration of USP39 rescues the suppressive effect on lung adenocarcinoma cell growth and migration induced by USP39 knockdown. A and B, Western blot assays were conducted to confirm the restoration of USP39 expression in both A549 and H1299 cell lines. Doxycycline at 1 µg/ml for 72 h were used to induce USP39 knockdown. Columns on the right show the quantification of relative levels of USP39. C, Colony formation assay performed using A549 or H1299 parental cells, A549 or H1299 cells stably expressing USP39-shRNA1 vector, and these cells together with USP39 rescue. Graphs show the quantification of relative colony sizes in each group. D and E, Cell proliferation were detected by EdU staining using A549 and H1299 cell lines in the condition of USP39 slicing or complementation. Representative images were exhibited. Scale bar, 200 μm. F, Graphs show the quantification of relative levels of EdU positive cells indicated in D and E. G, MTT assays were performed to detect cell viability in A549 or H1299 parental, USP39 knockdown and restoration cells. H, Representative images of transwell assays in A549 or H1299 parental cells and A549 or H1299 cells with USP39 knockdown and resto ration. Scale bar, 100 μm. I, Quantification data of cell numbers in H. Three independent repeats were conducted, with ** indicates p < 0.01

Figure Legend Snippet: Fig. 4 Restoration of USP39 rescues the suppressive effect on lung adenocarcinoma cell growth and migration induced by USP39 knockdown. A and B, Western blot assays were conducted to confirm the restoration of USP39 expression in both A549 and H1299 cell lines. Doxycycline at 1 µg/ml for 72 h were used to induce USP39 knockdown. Columns on the right show the quantification of relative levels of USP39. C, Colony formation assay performed using A549 or H1299 parental cells, A549 or H1299 cells stably expressing USP39-shRNA1 vector, and these cells together with USP39 rescue. Graphs show the quantification of relative colony sizes in each group. D and E, Cell proliferation were detected by EdU staining using A549 and H1299 cell lines in the condition of USP39 slicing or complementation. Representative images were exhibited. Scale bar, 200 μm. F, Graphs show the quantification of relative levels of EdU positive cells indicated in D and E. G, MTT assays were performed to detect cell viability in A549 or H1299 parental, USP39 knockdown and restoration cells. H, Representative images of transwell assays in A549 or H1299 parental cells and A549 or H1299 cells with USP39 knockdown and resto ration. Scale bar, 100 μm. I, Quantification data of cell numbers in H. Three independent repeats were conducted, with ** indicates p < 0.01

Techniques Used: Migration, Knockdown, Western Blot, Expressing, Colony Assay, Stable Transfection, Plasmid Preparation, Staining

Fig. 5 Transcriptomic profiling to identify downstream genes regulated by USP39. A, Venn diagram illustrates 719 overlapped DEGs in the H1299 parental (Ctrl) versus USP39 knockdown (USP39-sh1) and USP39 knockdown (USP39-sh1) versus USP39 rescue (USP39-sh1 + rescue) datasets. B, Heatmap exhibits the down- and up-regulation of the 719 DEGs in the comparison of indicated groups. C, Seven genes were selected with decreased expression in USP39 knockdown group and restored levels with USP39 complementation. D, RT-PCR were performed to test the expression alteration of selected genes with USP39 knockdown and restoration. E and F, Quantification data of the relative RNA levels of indicated genes in A549 (E) and H1299 (F) cell lines as detected in D. G, Western blot assays were performed to determine the changes of GLI1 expression at protein level in A549 and H1299 cells with USP39 knockdown and restoration. H, Quantification data showing the relative levels of GLI1 detected in G. Column charts represent data from 3 independent repeats. N.S. means no significance. * and ** indicate p < 0.05 and p < 0.01, respectively

Figure Legend Snippet: Fig. 5 Transcriptomic profiling to identify downstream genes regulated by USP39. A, Venn diagram illustrates 719 overlapped DEGs in the H1299 parental (Ctrl) versus USP39 knockdown (USP39-sh1) and USP39 knockdown (USP39-sh1) versus USP39 rescue (USP39-sh1 + rescue) datasets. B, Heatmap exhibits the down- and up-regulation of the 719 DEGs in the comparison of indicated groups. C, Seven genes were selected with decreased expression in USP39 knockdown group and restored levels with USP39 complementation. D, RT-PCR were performed to test the expression alteration of selected genes with USP39 knockdown and restoration. E and F, Quantification data of the relative RNA levels of indicated genes in A549 (E) and H1299 (F) cell lines as detected in D. G, Western blot assays were performed to determine the changes of GLI1 expression at protein level in A549 and H1299 cells with USP39 knockdown and restoration. H, Quantification data showing the relative levels of GLI1 detected in G. Column charts represent data from 3 independent repeats. N.S. means no significance. * and ** indicate p < 0.05 and p < 0.01, respectively

Techniques Used: Knockdown, Comparison, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Fig. 6 USP39 regulates GLI1 transcription and GLI1 knockdown inhibits cell growth and migration induced by USP39 overexpression. A, The luciferase activities were examined in HEK293T cells transfected with GLI1 firefly luciferase reporter and the plasmids of USP39 full length (FL), USP39-N(1-103), USP39-ΔN and catalytically inactive mutant USP39-C306A, respectively. The activities of renilla luciferase were measured as internal control. B, H1299 cells stably expressing Tet-on inducible USP39 shRNA1 and shRNA2 constructs were treated with doxycycline at 1 µg/ml for 72 h, and then transfected with GLI1 firefly luciferase reporter for 36 h. The luciferase activities were examined with renilla luciferase used as internal control. C, Western blot assays were performed to verify the efficiency of USP39 overexpression and GLI1 knockdown. D, MTT assays were conducted to detect the viability of A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. E, Colony formation assays were conducted in A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. F, Column charts show the quantification data of colony formation in E. G, Cell migration was investigated by transwell assays in A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. H, Column charts show the quantification of cell numbers. Three independent repeats were conducted, with * and ** indicate p < 0.05 and p < 0.01, respectively. N.S. means no significance

Figure Legend Snippet: Fig. 6 USP39 regulates GLI1 transcription and GLI1 knockdown inhibits cell growth and migration induced by USP39 overexpression. A, The luciferase activities were examined in HEK293T cells transfected with GLI1 firefly luciferase reporter and the plasmids of USP39 full length (FL), USP39-N(1-103), USP39-ΔN and catalytically inactive mutant USP39-C306A, respectively. The activities of renilla luciferase were measured as internal control. B, H1299 cells stably expressing Tet-on inducible USP39 shRNA1 and shRNA2 constructs were treated with doxycycline at 1 µg/ml for 72 h, and then transfected with GLI1 firefly luciferase reporter for 36 h. The luciferase activities were examined with renilla luciferase used as internal control. C, Western blot assays were performed to verify the efficiency of USP39 overexpression and GLI1 knockdown. D, MTT assays were conducted to detect the viability of A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. E, Colony formation assays were conducted in A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. F, Column charts show the quantification data of colony formation in E. G, Cell migration was investigated by transwell assays in A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. H, Column charts show the quantification of cell numbers. Three independent repeats were conducted, with * and ** indicate p < 0.05 and p < 0.01, respectively. N.S. means no significance

Techniques Used: Knockdown, Migration, Over Expression, Luciferase, Transfection, Mutagenesis, Control, Stable Transfection, Expressing, Construct, Western Blot

Fig. 7 USP39 slicing inhibits the growth of H1299 xenograft tumors. A, Mice were randomized to three group with implantation of H1299 control cells, H1299 stably transfected with USP39 shRNA, and H1299 stably transfected with USP39 shRNA together with rescue constructs, respectively. The wide and length of xenografts were measured every other day and the volumes were plotted. n = 6. B, Resected tumor weights were measured and plotted. C, Image exhibits the resected xenograft tumors from each group as indicated. D, Immunohistochemistry staining of USP39, Ki67 and GLI1 in tumor tissue sections from indicated group. Scale bar, 50 μm. Right column charts show quantification data of immunohistochemistry staining. E, Western blots were conducted to detect the protein levels of USP39 and GLI1 in lysed resected tumors. β-actin was used to confirm equal loading. Relative levels of GLI1 were quantified as shown on the right column charts. * and ** indicate p < 0.05 and p < 0.01, respectively

Figure Legend Snippet: Fig. 7 USP39 slicing inhibits the growth of H1299 xenograft tumors. A, Mice were randomized to three group with implantation of H1299 control cells, H1299 stably transfected with USP39 shRNA, and H1299 stably transfected with USP39 shRNA together with rescue constructs, respectively. The wide and length of xenografts were measured every other day and the volumes were plotted. n = 6. B, Resected tumor weights were measured and plotted. C, Image exhibits the resected xenograft tumors from each group as indicated. D, Immunohistochemistry staining of USP39, Ki67 and GLI1 in tumor tissue sections from indicated group. Scale bar, 50 μm. Right column charts show quantification data of immunohistochemistry staining. E, Western blots were conducted to detect the protein levels of USP39 and GLI1 in lysed resected tumors. β-actin was used to confirm equal loading. Relative levels of GLI1 were quantified as shown on the right column charts. * and ** indicate p < 0.05 and p < 0.01, respectively

Techniques Used: Control, Stable Transfection, Transfection, shRNA, Construct, Immunohistochemistry, Staining, Western Blot

Fig. 8 Graphic summary of USP39 phase separation in the nucleolus and its role in promoting lung adenocarcinoma progression. USP39 forms liquid- like condensates in the nucleolus via LLPS and IDR domain with residues 1-103 is indispensable for driving USP39 LLPS. USP39 condensates facilitate the oncogenic transcription factor GLI1 expression and lead to lung adenocarcinoma progression by promoting cell proliferation and migration. Conversely, downregulation of USP39 decreased GLI1 levels, and therefore inhibited cancer development

Figure Legend Snippet: Fig. 8 Graphic summary of USP39 phase separation in the nucleolus and its role in promoting lung adenocarcinoma progression. USP39 forms liquid- like condensates in the nucleolus via LLPS and IDR domain with residues 1-103 is indispensable for driving USP39 LLPS. USP39 condensates facilitate the oncogenic transcription factor GLI1 expression and lead to lung adenocarcinoma progression by promoting cell proliferation and migration. Conversely, downregulation of USP39 decreased GLI1 levels, and therefore inhibited cancer development

Techniques Used: Expressing, Migration

Incubation:

Article Title: USP39 facilitates breast cancer cell proliferation through stabilization of FOXM1
Article Snippet: .. Then the cells were incubated with mouse anti-FoxM1 (1:500, SantaCruz SC-376471) and rabbit anti-USP39 (1:300, Proteintech 23865-1-AP), followed by the incubation of a goat anti-rabbit IgG-FITC (1:500, Santa Cruz sc-2012) or goat anti-mouse IgG-PE (1:500, SantaCruz sc-3738) secondary antibody. .. The FluoView FV1000 confocal imaging system (Olympus) was used to image the cells.

Western Blot:

Article Title: USP39 facilitates breast cancer cell proliferation through stabilization of FOXM1
Article Snippet: .. The following antibodies and dilutions were used for Western blotting: rabbit anti-FOXM1 (1:1000, ABclonal, A2493), rabbit anti-FOXM1 (C-20) (1:5000, SantaCruz sc-502, recognizing FOXM1 C-terminus), rabbit anti-FOXM1 (K-19) (1:5000, SantaCruz sc-500, recognizing FOXM1 N-terminus), rabbit anti-USP39 (1:1000, Proteintech 23865-1-AP) mouse anti-flag (1:5000, Abmart # M22000 ), mouse anti-C-Myc (1:500, SantaCruz sc-40), mouse anti-GFP (1:500, Beyotime AG281), rabbit anti-GST (1:1000, Abcam ab19256), rabbit anti-CDC25B (1:1000, SantaCruz sc-326), rabbit anti-PLK1 (1:1000, CUSABIO, CSB- PA004668 ), mouse anti-Cdh1 (1:1000, SantaCruz sc-56312), rabbit anti-HA (1:1000, Proteintech 51064-2-AP), and mouse anti-β-actin (1:10000, Abcam ab49900). .. Signals from primary antibody were obtained via HRP conjugated anti-rabbit IgG (1:5000, ABclonal AS014) and anti-mouse IgG (1:5000; ABclonal AS003) magnification, and detected by the Kodak 4000 MM Imaging System using Super Signal West Femto Maximum Sensitivity Substrate (Thermo) (Kodak).

Image Search Results

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 1 USP39 is localized in nucleolus and undergoes LLPS. A, Representative images of immunofluorescence assay showing cellular localization of GFP-USP39 fusion protein expressed in HeLa cells with or without treatment of 3% 1, 6-hexanediol (1,6-HD) for 7 min. B, Analysis of intrinsic disorder tendency of USP39 using PhaSepDB database. Below schematic represents corresponding domains of USP39. C, FRAP analysis in HeLa cells transfected with GFP-USP39 showing the recovery of the condensate after photobleaching. D, Schematic illustration of sub-nucleolar compartments, including the fibrillar center (FC), dense fibrillar component (DFC), and granular component (GC), with indicated markers being RPA40, FBL and NPM1 respectively. E, Immunofluorescence analysis of colocalization of GFP-tagged USP39 with NPM1, RAP40, and FBL in HeLa cells. Diagram curves on the right show quan tification. Scale bar, 10 μm

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Immunofluorescence, Transfection

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 2 USP39 IDR domain is required for LLPS. A, Schematic representation of USP39 and its truncated mutants containing or deleting IDR domain. B, Representative images of immunofluorescence assay showing the localization of GFP-tagged USP39 with full length and the 5 truncated versions of GFP- USP39 in HeLa cells. C, FRAP assay in HeLa cells transfected with GFP-USP39-N(1-103) showing the recovery of bleached droplet foci. D, Representative images of RC and fluorescence microscopy showing in vitro droplet formation of purified GFP vector and GFP-USP39-N(1-103) fusion protein both at 60 µM in the presence of PEG 4000 and 150 mM of NaCl. E, Representative images of RC and fluorescence microscopy showing in vitro droplet formation of purified GFP-USP39-N(1-103) fusion protein (60 µM) with escalating molecular weights of the PEG crowders as indicated. Scale bar, 10 μm

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Immunofluorescence, FRAP Assay, Transfection, Fluorescence, Microscopy, In Vitro, Purification, Plasmid Preparation

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 3 The definite condition of USP39 droplet formation was detected in serial concentration gradients. A and B, Diagram showing in vitro droplet formation of purified GFP-USP39-N(1-103) in the absence (A) or presence (B) of PEG 4000 with NaCl concentrations at 150, 200, 250, and 300 mM. The concentration of fusion protein added in buffer were as indicated. C, Representative images exhibiting droplet formation of GFP-USP39-N(1-103) fusion protein at the concentrations of 3, 6, 12, 20, 30 and 60 µM with NaCl concentrations ranging from 150 to 300 mM as illustrated in b. Scale bar, 10 μm

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Concentration Assay, In Vitro, Purification

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 4 Restoration of USP39 rescues the suppressive effect on lung adenocarcinoma cell growth and migration induced by USP39 knockdown. A and B, Western blot assays were conducted to confirm the restoration of USP39 expression in both A549 and H1299 cell lines. Doxycycline at 1 µg/ml for 72 h were used to induce USP39 knockdown. Columns on the right show the quantification of relative levels of USP39. C, Colony formation assay performed using A549 or H1299 parental cells, A549 or H1299 cells stably expressing USP39-shRNA1 vector, and these cells together with USP39 rescue. Graphs show the quantification of relative colony sizes in each group. D and E, Cell proliferation were detected by EdU staining using A549 and H1299 cell lines in the condition of USP39 slicing or complementation. Representative images were exhibited. Scale bar, 200 μm. F, Graphs show the quantification of relative levels of EdU positive cells indicated in D and E. G, MTT assays were performed to detect cell viability in A549 or H1299 parental, USP39 knockdown and restoration cells. H, Representative images of transwell assays in A549 or H1299 parental cells and A549 or H1299 cells with USP39 knockdown and resto ration. Scale bar, 100 μm. I, Quantification data of cell numbers in H. Three independent repeats were conducted, with ** indicates p < 0.01

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Migration, Knockdown, Western Blot, Expressing, Colony Assay, Stable Transfection, Plasmid Preparation, Staining

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 5 Transcriptomic profiling to identify downstream genes regulated by USP39. A, Venn diagram illustrates 719 overlapped DEGs in the H1299 parental (Ctrl) versus USP39 knockdown (USP39-sh1) and USP39 knockdown (USP39-sh1) versus USP39 rescue (USP39-sh1 + rescue) datasets. B, Heatmap exhibits the down- and up-regulation of the 719 DEGs in the comparison of indicated groups. C, Seven genes were selected with decreased expression in USP39 knockdown group and restored levels with USP39 complementation. D, RT-PCR were performed to test the expression alteration of selected genes with USP39 knockdown and restoration. E and F, Quantification data of the relative RNA levels of indicated genes in A549 (E) and H1299 (F) cell lines as detected in D. G, Western blot assays were performed to determine the changes of GLI1 expression at protein level in A549 and H1299 cells with USP39 knockdown and restoration. H, Quantification data showing the relative levels of GLI1 detected in G. Column charts represent data from 3 independent repeats. N.S. means no significance. * and ** indicate p < 0.05 and p < 0.01, respectively

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Knockdown, Comparison, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 6 USP39 regulates GLI1 transcription and GLI1 knockdown inhibits cell growth and migration induced by USP39 overexpression. A, The luciferase activities were examined in HEK293T cells transfected with GLI1 firefly luciferase reporter and the plasmids of USP39 full length (FL), USP39-N(1-103), USP39-ΔN and catalytically inactive mutant USP39-C306A, respectively. The activities of renilla luciferase were measured as internal control. B, H1299 cells stably expressing Tet-on inducible USP39 shRNA1 and shRNA2 constructs were treated with doxycycline at 1 µg/ml for 72 h, and then transfected with GLI1 firefly luciferase reporter for 36 h. The luciferase activities were examined with renilla luciferase used as internal control. C, Western blot assays were performed to verify the efficiency of USP39 overexpression and GLI1 knockdown. D, MTT assays were conducted to detect the viability of A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. E, Colony formation assays were conducted in A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. F, Column charts show the quantification data of colony formation in E. G, Cell migration was investigated by transwell assays in A549 and H1299 cell lines stably transfected with pCDH, pCDH-USP39, and pCDH-USP39 together with shGLI1 constructs. H, Column charts show the quantification of cell numbers. Three independent repeats were conducted, with * and ** indicate p < 0.05 and p < 0.01, respectively. N.S. means no significance

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Knockdown, Migration, Over Expression, Luciferase, Transfection, Mutagenesis, Control, Stable Transfection, Expressing, Construct, Western Blot

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 7 USP39 slicing inhibits the growth of H1299 xenograft tumors. A, Mice were randomized to three group with implantation of H1299 control cells, H1299 stably transfected with USP39 shRNA, and H1299 stably transfected with USP39 shRNA together with rescue constructs, respectively. The wide and length of xenografts were measured every other day and the volumes were plotted. n = 6. B, Resected tumor weights were measured and plotted. C, Image exhibits the resected xenograft tumors from each group as indicated. D, Immunohistochemistry staining of USP39, Ki67 and GLI1 in tumor tissue sections from indicated group. Scale bar, 50 μm. Right column charts show quantification data of immunohistochemistry staining. E, Western blots were conducted to detect the protein levels of USP39 and GLI1 in lysed resected tumors. β-actin was used to confirm equal loading. Relative levels of GLI1 were quantified as shown on the right column charts. * and ** indicate p < 0.05 and p < 0.01, respectively

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Control, Stable Transfection, Transfection, shRNA, Construct, Immunohistochemistry, Staining, Western Blot

Journal: Cell communication and signaling : CCS

Article Title: USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression.

doi: 10.1186/s12964-025-02059-5

Figure Lengend Snippet: Fig. 8 Graphic summary of USP39 phase separation in the nucleolus and its role in promoting lung adenocarcinoma progression. USP39 forms liquid- like condensates in the nucleolus via LLPS and IDR domain with residues 1-103 is indispensable for driving USP39 LLPS. USP39 condensates facilitate the oncogenic transcription factor GLI1 expression and lead to lung adenocarcinoma progression by promoting cell proliferation and migration. Conversely, downregulation of USP39 decreased GLI1 levels, and therefore inhibited cancer development

Article Snippet: For Western blot and immunohistochemistry assays, rabbit anti-USP39 antibody (Proteintech, 23865-1-AP) and mouse anti-GLI1 antibody (Proteintech, 66905-1-Ig) were used.

Techniques: Expressing, Migration

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rabbit anti usp39 antibody (Proteintech)

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1) Product Images from "USP39 phase separates into the nucleolus and drives lung adenocarcinoma progression by promoting GLI1 expression."

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