Journal: The EMBO Journal

Article Title: HDAC6-dependent deacetylation of SAE2 enhances SUMO1 conjugation for mitotic integrity

doi: 10.1038/s44318-025-00532-y

Figure Lengend Snippet: ( A ) Western blot, representative of three, of acetyl-K164-SAE2 (mouse monoclonal) following anti-FLAG immunoprecipitation from U2OS cells and U2OS cells expressing FLAG-SAE2 in unsynchronised cells or cells treated with nocodazole for 17 h before washing and releasing into mitosis for 10 min. HDAC inhibitors, Panobinostat (HDAC class I, II, and IV), ACY-738 (HDAC6), and RGFP966 (HDAC3), were applied to cells in the last 2 h of nocodazole synchronisation at 2.5 µM and reapplied upon the release. An antibody specific for phosphorylated-Ser10 on histone 3 is used as a marker of mitosis. Quantification of the relative abundance of acetyl-K164-SAE2 relative to the total abundance of SAE2 immunoprecipitated. Error bars = SEM; N = 3 biological repeats. *** P < 0.001, ** P < 0.01, * P < 0.05, ns = not significant P > 0.05. Vehicle vs Nocodazole P = 0.0002, Nocodazole vs Pan-HDACi P = 0.0143, Nocodazole vs HDAC6i P = 0.0019, Nocodazole vs HDAC3i P = 0.9494. Statistical significance was calculated using one-way ANOVA. ( B ) Western blot analysis of a FLAG-SAE2 co-immunoprecipitation with HDAC6 from U2OS, in the context of an asynchronous cell population or following a 16 h nocodazole-treatment and mitotic shake-off. Replicated twice in the laboratory. ( C ) Western blot of acK164-SAE2 following anti-FLAG immunoprecipitation from U2OS cells and U2OS cells expressing FLAG-SAE2 in asynchronous cells treated with indicated combinations of inhibitors against the histone acetyltransferases p300 (A-485), TIP60 (NU9056), NAT10 (Remodelin Hydrobromide), and GCN5 (Butyrolactone). Inhibitors were added for 2 h at 2.5 µM. Replicated three times in the laboratory.

Article Snippet: Panobinostat , SignalChem , H83-904G.

Techniques: Western Blot, Immunoprecipitation, Expressing, Marker

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Panobinostat increases histone H3 (Lys9) and H4 (Lys8) acetylation in TNBC cell lines . Cells were treated with panobinostat (100, 200 nM) or vehicle (DMSO) for 18 hours, fixed, permeabilized and stained for acetyl-histones (A) H3 (Lys9) or (B) H4 (Lys8) and subjected to flow cytometry. Data are presented as mean fluorescence intensity (mean ± SEM) of two independent experiments, (***, P < 0.001). (C-D) Confocal images of TNBC cell lines treated with panobinostat (100 nM) or vehicle for 18 hours, fixed, permeabilized and stained red (rhodamine phalloidin) for actin filaments and green (Alexa Fluor ® 488) for acetyl-histones (C) H3 (Lys9) or (D) H4 (Lys8). Original magnification was 400× with scale bars at 20 microns. DMSO, dimethyl sulfoxide; SEM, standard error of the mean; TNBC, triple-negative breast cancer.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques: Staining, Flow Cytometry, Fluorescence

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Panobinostat decreases TNBC cell proliferation and viability . Cells from four TNBC cell lines (MDA-MB-157, MDA-MB-231, MDA-MB-468, BT549) and three ER-positive cell lines (MCF-7, MDA-MB-361, ZR-75) were treated with panobinostat (50, 100, 200 nM) or vehicle (DMSO) for 24 hours and assayed by (A) MTT proliferation and (B) trypan blue exclusion assays. Data are represented as percent control (mean ± SEM) of three independent experiments, (**, P < 0.01; ***, P < 0.001). DMSO, dimethyl sulfoxide; ER, estrogen receptor; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; SEM, standard error of the mean; TNBC, triple-negative breast cancer.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques: Control

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Effect of 100 nM panobinostat on cell cycle percentage of TNBC cells.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Panobinostat induces apoptosis in TNBC cells . (A) TNBC cells treated with panobinostat (100, 200 nM) or vehicle (DMSO) for 24 hours were assayed by DNA fragmentation (Cell Death ELISA) assay to assess changes in apoptosis. Data are presented as enrichment (mean ± SEM) versus control of two independent experiments (***, P < 0.001). (B) Representative confocal images show the presence of apoptotic bodies (arrows) in panobinostat treated MDA-MB-157, MDA-MB-231, and BT-549 cells at 18 hours. Cells stained red (rhodamine phalloidin) for actin filaments, green (Alexa Fluor ® 488) for acetyl-histone H3 (Lys9), and blue for DAPI nuclear stain. Original magnification is 400× with scale bars at 20 microns. DAPI, 4',6-diamidino-2-phenylindole; DMSO, dimethyl sulfoxide; SEM, standard error of the mean; TNBC, triple-negative breast cancer.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Staining

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Effect of panobinostat on tumor growth in MDA-MB-231 and BT549 xenograft models . Female, CB-17/SCID mice ( n = 5/group) were injected with (A) MDA-MB-231-tRFP or (B) BT-549-tRFP cells (5 × 10 6 cells/injection) bilaterally into the inguinal mammary fat pad. On day 14, mice were treated intraperitoneally (i.p.) with panobinostat (10 mg/kg) or vehicle (1:20 DMSO in normal saline) five days/week for 28 days. Data points represent average tumor volume ± SEM, (***, P < 0.001). DMSO, dimethyl sulfoxide; SEM, standard error of the mean; tRFP, turbo red fluorescent protein.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques: Injection, Saline

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Heat map of panobinostat -induced gene expression changes in MDA-MB-231, MDA-MB-468, and MCF-7 cells . MDA-MB-231, MDA-MB-468, or MCF-7 cells were treated for 24 hours with panobinostat (100 nM) or vehicle (DMSO) and then assayed via the Human Breast Cancer and Estrogen Receptor Signaling RT 2 Profiler™ PCR Array. Red signifies up-regulation and green signifies down-regulation by panobinostat compared to MDA-MB-231 vehicle treated controls. Data are representative of three independent experiments. Genes regulated at least 2-fold are also summarized in Additional file (MDA-MB-231), 2 (MDA-MB-468) and 3 (MCF-7). DMSO, dimethyl sulfoxide.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques: Gene Expression

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Panobinostat up-regulates CDH1 expression and initiates EMT reversal in MDA-MB-231 Cells . MDA-MB-231 cells were plated overnight and treated with panobinostat (100 nM) or vehicle (DMSO) for 24 hours. The expression of CDH1 was examined by (A) flow cytometry and (B) ELISA. Data are represented as mean ± SEM of two independent experiments, (***, P < 0.001). (C-E) MDA-MB-231 morphology changes were assessed in vehicle- and panobinostat- (100 n M) treated cells with three-color fluorescence staining on a BD Pathway 855 Bioimager. (C) Control and (D-E) Panobinostat treated cells were stained red (rhodamine phalloidin) for actin filaments, green (Alexa Fluor ® 488) for acetyl-histone H3 (Lys9), and blue (DAPI) nuclear counter stain. Partial reversal of EMT in treated cells is indicated by the presence of cuboidal/spherical cells (arrows). (E) Two-fold magnification of field with normal untransformed mesenchymal cell and transformed spherical cells. Original magnification is 400× with scale bars at 20 microns. DAPI, 4',6-diamidino-2-phenylindole; DMSO, dimethyl sulfoxide; ELISA, enzyme-linked immunosorbent assay; EMT, epithelial-to-mesenchymal transition; SEM, standard error of the mean.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques: Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Fluorescence, Staining, Control, Transformation Assay

Journal: Breast Cancer Research : BCR

Article Title: Targeting triple-negative breast cancer cells with the histone deacetylase inhibitor panobinostat

doi: 10.1186/bcr3192

Figure Lengend Snippet: Panobinostat increases CDH1 expression in MDA-MB-231 primary tumors . Control and panobinostat treated, formalin-fixed mammary fat pad sections from MDA-MB-231-tRFP injected CB17-SCID mice were stained for H & E (left column) or anti-human CDH1 (1:30; right column) followed by Alexa Fluor ® 488 secondary antibody. Original magnification was 100× with scale bars at 200 microns. tRFP, turbo red fluorescent protein; SCID, severe combined immunodeficiency.

Article Snippet: Panobinostat was dissolved in dimethyl sulfoxide (DMSO) (Invitrogen) as a 1 mM stock solution and kept at -20°C.

Techniques: Expressing, Control, Injection, Staining

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Inhibitors of Signaling Pathways That Block Reversal of HIV-1 Latency

doi: 10.1128/AAC.01744-18

Figure Lengend Snippet: Kinase inhibitors identified in the screen that blocked the latency-reversing activity of prostratin, panobinostat, or JQ-1 a

Article Snippet: Prostratin, panobinostat, and JQ1 were acquired from Sigma (St. Louis, MO), Selleckchem, and BPS Bioscience (San Diego, CA), respectively.

Techniques: Activity Assay, Inhibition

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Inhibitors of Signaling Pathways That Block Reversal of HIV-1 Latency

doi: 10.1128/AAC.01744-18

Figure Lengend Snippet: Characterization of PF-3758309, danusertib, AZ628, and P276-00 in 24ST1NLESG cells a , b

Article Snippet: Prostratin, panobinostat, and JQ1 were acquired from Sigma (St. Louis, MO), Selleckchem, and BPS Bioscience (San Diego, CA), respectively.

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Inhibitors of Signaling Pathways That Block Reversal of HIV-1 Latency

doi: 10.1128/AAC.01744-18

Figure Lengend Snippet: Characterization of CDK inhibitors in the in 24ST1NLESG cells a

Article Snippet: Prostratin, panobinostat, and JQ1 were acquired from Sigma (St. Louis, MO), Selleckchem, and BPS Bioscience (San Diego, CA), respectively.

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Inhibitors of Signaling Pathways That Block Reversal of HIV-1 Latency

doi: 10.1128/AAC.01744-18

Figure Lengend Snippet: Characterization of mTOR inhibitors in the in 24ST1NLESG cells a

Article Snippet: Prostratin, panobinostat, and JQ1 were acquired from Sigma (St. Louis, MO), Selleckchem, and BPS Bioscience (San Diego, CA), respectively.

Techniques: