Journal: Frontiers in Cell and Developmental Biology

Article Title: Transcriptional Repression of Ferritin Light Chain Increases Ferroptosis Sensitivity in Lung Adenocarcinoma

doi: 10.3389/fcell.2021.719187

Figure Lengend Snippet: KAT5-induced FTL acetylation suppresses ferroptosis. (A,B) STRING analysis revealed TFCP2 interacted acyltransferase (A) and methyltransferases (B) (confidence = 0.4). (C) Co-immunoprecipitation experiments performed in PC9 cells using IgG, anti-KAT5 and anti-TFCP2 antibodies, and further analysis of KAT5, TFCP2 and GAPDH expression by IB. (D–G) The enrichments of H4K12Ac and H4K16Ac at –2k, TFCP2 (FOXA1) motif or 2k regions of FTL (D,F) or FTH (E,G) promoter were calculated as the percentage of input chromosomal DNA via ChIP using the corresponding antibodies in WT (D,E) or KAT5 –/– (F, G) PC9 cells with erastin (10 μM) treated for indicated times. IgG was used as the parallel control. (H) FTH expression were measured using IB in WT, YAP –/– , TFCP2 –/– and KAT5 –/– PC9 cells with erastin (10 μM) treatment for indicated hours. (I–K) Labile iron, cell death and lipid ROS were measured in PC9 cells with or without KAT5 overexpression combined with FTL knockdown before erastin (10 μM, 24 h or 16 h) treatment. (L,M) Labile iron, cell death and lipid ROS generation were measured in PC9 and H1299 cells with or without FTL knocked down. Pentamidine (30 μM) and TH1834 (100 μM) were used to pretreat cells for 8 h before further treated with erastin (10 μM, 24 h). (N) The enrichments of YAP, TFCP2 and FOXA1 at –2k, TFCP2 (FOXA1) motif or 2k regions of FTL promoter were calculated as the percentage of input chromosomal DNA via ChIP using the corresponding antibodies in PC9 cells with or without KAT5 overexpressed. (O) Enrichment of YAP at TFCP2 (FOXA1)-binding motif within the FTL promoter in WT or KAT5 –/– PC9 cells with or without YAP, TFCP2 and FOXA1 overexpressed was measured by ChIP-qPCR. (P) Co-IP experiments were performed using anti-YAP in PC9 cells with KAT5 overexpression or knockdown with or without erastin treatment (10 μM, 24 h). The YAP level in each co-IP samples was adjusted to the same protein content. The indicated proteins in co-IP samples or WCL were measured by IB. The data are shown as the mean ± SD from three biological replicates (including IB). * P < 0.05, ** P < 0.01 indicates statistical significance. Data in (I–M,O) were analyzed using a one-way ANOVA test. Data in (N) were analyzed using student’s t -test.

Article Snippet: Cells were treated with erastin (Selleckchem, Houston, TX, United States), cycloheximide (CHX, Sigma, St Louis, MO, United States), cisplatin (MedChemExpress, Monmouth Junction, NJ, United States), AZD9291 (MedChemExpress), pentamidine (MedChemExpress), TH1834 (Sigma), Dox (Clontech, Mountain View, CA, United States), EGCG (MedChemExpress), EGTA (MedChemExpress), H89 (MedChemExpress), Rp-cAMPs (MedChemExpress), glucose (Sigma), GlcNAc (Sigma), GlcN (MedChemExpress), DFO (Selleckchem), or CPX (Selleckchem).

Techniques: Immunoprecipitation, Expressing, Control, Over Expression, Knockdown, Binding Assay, Co-Immunoprecipitation Assay

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Pentamidine Inhibits Titanium Particle-Induced Osteolysis In Vivo and Receptor Activator of Nuclear Factor-κB Ligand-Mediated Osteoclast Differentiation In Vitro

doi: 10.1007/s13770-019-00186-y

Figure Lengend Snippet: Pentamidine prevented bone erosion in Ti particle-induced osteolysis in a mouse calvarial model. Ti particles were embedded on to the mouse calvaria, and injected with pentamidine (1 or 5 mg/kg) or vehicle control for 10 days. At the end of the study period, calvaria were dissected, fixed, and scanned with micro-CT. A Representative micro-CT images of calvaria from each group (n = 5). Scale bar = 5 mm. B Quantification of bone mineral density (BMD) and bone volume to tissue volume (BV/TV) ratio. **p < 0.01 and *p < 0.05. Pent: pentamidine

Article Snippet: To induce osteoclast differentiation, BMMs were cultured in osteoclastogenic medium (20 ng/mL RANKL and 10 ng/mL M-CSF) in the presence or absence of different doses of pentamidine isethionate (Selleck Chemicals, Houston, TX, USA).

Techniques: Injection, Control, Micro-CT

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Pentamidine Inhibits Titanium Particle-Induced Osteolysis In Vivo and Receptor Activator of Nuclear Factor-κB Ligand-Mediated Osteoclast Differentiation In Vitro

doi: 10.1007/s13770-019-00186-y

Figure Lengend Snippet: Histological analysis of murine calvarial bone sections. A Representative images of hematoxylin and eosin (H&E), and tartrate-resistant acid phosphatase (TRAP) stained calvarial bone sections. Scale bar = 100 μm. B The number of TRAP-positive cells (yellow arrow head) per bone surface in each group (n = 5) was assessed (lower graph). *p < 0.05 versus Ti group. Pent: pentamidine

Article Snippet: To induce osteoclast differentiation, BMMs were cultured in osteoclastogenic medium (20 ng/mL RANKL and 10 ng/mL M-CSF) in the presence or absence of different doses of pentamidine isethionate (Selleck Chemicals, Houston, TX, USA).

Techniques: Staining

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Pentamidine Inhibits Titanium Particle-Induced Osteolysis In Vivo and Receptor Activator of Nuclear Factor-κB Ligand-Mediated Osteoclast Differentiation In Vitro

doi: 10.1007/s13770-019-00186-y

Figure Lengend Snippet: Pentamidine suppressed RANKL-induced osteoclast differentiation in vitro and the expression of osteoclast-specific genes. A Bone marrow-derived macrophages (BMMs) were cultured in osteoclast-inducing medium containing M-CSF (10 ng/mL) and RANKL (20 ng/mL) in the presence or absence of various concentrations of pentamidine. After 4 days of culture, cells were fixed and stained for TRAP. Scale bar = 500 μm. B The number of TRAP-positive multinucleated cells was counted. C BMMs were treated with different doses of pentamidine in the presence of M-CSF for 3 days. Cell viability was determined by using MTT assay. D BMMs were incubated with M-CSF (10 ng/mL) and RANKL (20 ng/mL) in the presence or absence of 5 μM pentamidine for 4 days and mRNA levels of NFATc1 (Nfatc1), Cathepsin K (Ctsk), DC-STAMP (Dcstamp), and TRAP (Acp5) were determined by real-time PCR. **p < 0.01 and *p < 0.05 versus vehicle-treated control group

Article Snippet: To induce osteoclast differentiation, BMMs were cultured in osteoclastogenic medium (20 ng/mL RANKL and 10 ng/mL M-CSF) in the presence or absence of different doses of pentamidine isethionate (Selleck Chemicals, Houston, TX, USA).

Techniques: In Vitro, Expressing, Derivative Assay, Cell Culture, Staining, MTT Assay, Incubation, Real-time Polymerase Chain Reaction, Control

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Pentamidine Inhibits Titanium Particle-Induced Osteolysis In Vivo and Receptor Activator of Nuclear Factor-κB Ligand-Mediated Osteoclast Differentiation In Vitro

doi: 10.1007/s13770-019-00186-y

Figure Lengend Snippet: Pentamidine inhibited actin ring formation and nuclear localization of NFATc1, and impaired RANKL-induced NF-κB signaling. A BMMs were seeded on glass coverslips and stimulated with M-CSF (10 ng/mL) and RANKL (20 ng/mL) in the presence or absence of 5 μM pentamidine. Nuclear localization of NFATc1 was examined by immunostaining. Nuclei and F-actin were labeled with DAPI and rhodamine-conjugated phalloidin, respectively. Scale bar = 50 μm. B Serum-starved BMMs were pretreated with or without 5 μM pentamidine for 1 h followed by stimulation with RANKL (50 ng/mL). Western blot analysis was performed to determine the levels of phosphorylated p38 (p-p38), p-ERK, p-JNK, and p-IκBα by using specific antibodies

Article Snippet: To induce osteoclast differentiation, BMMs were cultured in osteoclastogenic medium (20 ng/mL RANKL and 10 ng/mL M-CSF) in the presence or absence of different doses of pentamidine isethionate (Selleck Chemicals, Houston, TX, USA).

Techniques: Immunostaining, Labeling, Western Blot

Journal: Physical chemistry chemical physics : PCCP

Article Title: Glycosaminoglycans are potential pharmacological targets for classic DNA minor groove binder drugs berenil and pentamidine.

doi: 10.1039/c5cp03153b

Figure Lengend Snippet: Scheme 1 Chemical structures of the cationic form of berenil (top) and pentamidine.

Article Snippet: Berenil (diminazene aceturate, cat# sc-205651, purity 97.5%) and pentamidine isethionate (cat# sc-204176, purity Z 99%) were obtained from Santa Cruz Biotechnology, Inc. Heparin sodium salt from porcine intestinal mucosa (Sigma, cat# 51551) and chondroitin 6-sulfate sodium salt from shark cartilage (Sigma, cat# C4384) were used as supplied.

Techniques:

Journal: Physical chemistry chemical physics : PCCP

Article Title: Glycosaminoglycans are potential pharmacological targets for classic DNA minor groove binder drugs berenil and pentamidine.

doi: 10.1039/c5cp03153b

Figure Lengend Snippet: Fig. 4 Induced CD and absorption spectra of pentamidine measured at increasing concentrations of CS in deionized water (25 1C). Molar absorp- tion (e) and circular dichroic absorption coefficients (De) calculated by using drug concentration are shown in parentheses. Arrows denote isosbestic points at 231 and 280 nm.

Article Snippet: Berenil (diminazene aceturate, cat# sc-205651, purity 97.5%) and pentamidine isethionate (cat# sc-204176, purity Z 99%) were obtained from Santa Cruz Biotechnology, Inc. Heparin sodium salt from porcine intestinal mucosa (Sigma, cat# 51551) and chondroitin 6-sulfate sodium salt from shark cartilage (Sigma, cat# C4384) were used as supplied.

Techniques: Concentration Assay

Journal: Physical chemistry chemical physics : PCCP

Article Title: Glycosaminoglycans are potential pharmacological targets for classic DNA minor groove binder drugs berenil and pentamidine.

doi: 10.1039/c5cp03153b

Figure Lengend Snippet: Fig. 5 Induced CD and absorption spectra of pentamidine measured at increasing concentrations of heparin (50 mm phosphate buffer at pH 7.0, 80 mm Na+, 25 1C). Molar absorption (e) and circular dichroic absorption coefficients (De) calculated by using drug concentration are shown in parentheses. Thin line shows the smoothed ICD curve of PNT measured at 246 mM heparin. Arrow denotes an isosbestic point at 285 nm.

Article Snippet: Berenil (diminazene aceturate, cat# sc-205651, purity 97.5%) and pentamidine isethionate (cat# sc-204176, purity Z 99%) were obtained from Santa Cruz Biotechnology, Inc. Heparin sodium salt from porcine intestinal mucosa (Sigma, cat# 51551) and chondroitin 6-sulfate sodium salt from shark cartilage (Sigma, cat# C4384) were used as supplied.

Techniques: Concentration Assay