Journal: bioRxiv

Article Title: Arsenite methyltransferase 3 regulates hepatic energy metabolism which dictates the hepatic response to arsenic exposure

doi: 10.1101/2023.04.05.535637

Figure Lengend Snippet: A . Representative images of WT and as3mt -/- larvae and adults. B-C . Standard length of larvae (μM; B) and adults (cm; C). D . Average IC-ICP-MS counts of AsB, AsIII, DMA, and MMA from WT (grey) or as3mt -/- (pink) zebrafish extracts after 0 (dotted lines) or 1 mM iAs (6-120 hpf) challenge (solid lines) (n = 40, 2 clutches). E . Percent survival of WT (grey) and as3mt -/- (pink) across 0, 0.5, 1.0, 1.5 and 2.0 mM iAs exposure from 6-120 hpf. Intersect with green line denotes the LC 50 . n = ∼60, 3 clutches. F . Representative images of WT and as3mt -/- larvae during 0 mM, 1 mM acute (96-120 hpf), or 1 mM chronic (6-120 hpf) exposure to iAs. G . Images of 120 hpf zebrafish pigmentation at 0 mM or 1 mM chronic (6-120 hpf) iAs challenge (purple). Orange arrows mark pigmentation clusters. H . Quantification of pigmentation per larvae (n = 5-10, 1 clutch). I . Percent phenotype (normal (white), abnormal (grey) or dead (black) after 0-2 mM iAs exposure from 6-120 hpf (n = ∼60, 3 clutches).

Article Snippet: IC-ICP-MS standards were: As(III) (Arsenic (III) Standard, Fisher Scientific; 1327-53-3), As(V) (Arsenic V Speciation Standard, Fisher Scientfic; 7732-18-5, AsB (Arsenobetaine Standard Solution, LGC Standards; NIST-3033), DMA (Dimethylarsinic Acid Standard Solution, LGC Standards; NIST-3031) and MMA (Monomethylarsonic Acid Standard Solution, LGC Standards; NIST-3030).

Techniques:

Journal: Journal of Biological Chemistry

Article Title: Atypical Protein Kinase Cι Plays a Critical Role in Human Lung Cancer Cell Growth and Tumorigenicity

doi: 10.1074/jbc.m505402200

Figure Lengend Snippet: FIG. 2. PKC does not regulate NFB signaling in A549 cells. A, immunoblot analysis of A549 cell transfectants for cIAP2, Bcl-XL, and actin. B, analysis of A549 cell transfectants for PARP and cleaved PARP. HeLa cells treated with taxol for either 24 or 48 h served as a positive control for cleaved PARP. C, A549/pBabe and A549/kdPKC cells were treated with 50 g/ml TNF for the indicated times and cell lysates subjected to immunoblot analysis for detection of IB, phospho-(Ser32)-IB, and actin. D, A549/pBabe and A549/kdPKC cells were treated with 50 g/ml TNF for the indicated times and cell lysates subjected to immunoblot analysis for detection of NFB p65, phospho-(Ser536)-NFB, p65, and actin. E, transcriptional activity of an NFB-luciferase reporter in A549/pBabe and A549/kdPKC cells in the presence and absence of TNF. Data represent the mean S.E. of three independent determinations.

Article Snippet: This paper is available on line at http://www.jbc.org 31109 at K U N G L IG A T E K N ISK A H O G SK O L A N on M arch 8, 2015 http://w w w .jbc.org/ D ow nloaded from (Sigma, catalog number A8592); Rac1, 1:3000 (BD Transduction Laboratories, catalog number 610651); cIAP2, 1:500 (Santa Cruz Biotechnology, catalog number sc-7944); Bcl-XL, 1:1,000 (Cell Signaling Technologies, catalog number 2762); PARP/cleaved PARP, 1:1,000 (Cell Signaling Technologies, catalog number 9542); I B , 1:1,000 (Cell Signaling Technologies, catalog number 9242); phospho-(Ser32)-I B , 1:1,000 (Cell Signaling Technologies, catalog number 9241); NF B p65, 1:1,000 (Cell Signaling Technologies, catalog number 3034); phospho(Ser536)-NF B p65, 1:1,000 (Cell Signaling Technologies, catalog number 3031); Mek, phospho-(Ser217/221)-Mek, and phospho-(Ser298)Mek, each at 1:1,000 (Cell Signaling Technologies, catalog numbers 9122, 9121, and 9128, respectively), Erk and phospho-(Thr202/Tyr204)Erk, each at 1:1,000 (Cell Signaling Technologies, catalog numbers 9102 and 9101, respectively); CD31, 1:1,000 (or Pecam-1; Santa Cruz Biotechnology, catalog number sc-1506); BrdUrd, 1:100 (DAKO, catalog number M0744).

Techniques: Western Blot, Positive Control, Activity Assay, Luciferase

Journal: Cancer Research

Article Title: SAICAR Drives T Regulatory Cell Differentiation and FOXP3 Maintenance to Promote Immunotherapy Resistance

doi: 10.1158/0008-5472.CAN-25-4373

Figure Lengend Snippet: SAICAR promotes TGFβ–SMAD3 signaling by binding PPM1A. A, Summary of gene expression related to TGFβ–SMAD3 signaling and Jak–STAT5 signaling from RNA-seq of naïve CD4 + T from WT mice, which were cultured under Treg differentiation with or without SAICAR for 48 hours ( n = 3). B and C, Statistics showing Foxp3 mRNA expression by qPCR of naïve CD4 + T cells sorted from Adsl f/f - Cd4 Cre or Adsl f/f ( B ) and Paics f/f - Cd4 Cre or Paics f/f ( C ) mice under Treg differentiation conditions. Experiments were repeated four times. D and E, ChIP assay to measure the combination of SMAD3 and Foxp3 enhancer in naïve CD4 + T cells from Adsl f/f - Cd4 Cre or Adsl f/f ( D ) and Paics f/f - Cd4 Cre or Paics f/f ( E ) mice under Treg differentiation. Experiments were repeated four times. Homologous IgG and naïve CD4 T cells from Adsl f/f ( D ) or Paics f/f ( E ) mice were used for negative control. F, Statistics showing the percentage of Th17 cells in CD4 + T cells after naïve CD4 + T cells were present in Th17 differentiation conditions with or without SAICAR. Experiments were repeated five times. G, Statistics showing the percentage of Treg cells in CD4 + T cell of naïve CD4 + T cells, which were present in Treg differentiation conditions under SIS (SMAD3 inhibitor) and STAT5-IN-1 (STAT5 inhibitor) with or without SAICAR. Experiments were repeated three times. H, Naïve CD4 + T cells isolated from indicated mice, which were present in Treg differentiation conditions, and the percentage of Treg cells in CD4 + T cells was analyzed. Experiments were repeated three times. I and J, MC38 tumor cells were implanted into indicated mice, and the tumor growth curves ( n = 5; I ) and the percentage of Treg cells in CD4 + T cells are shown ( n = 5; J ). K, Whole-cell extracts (WCE) of naïve CD4 + T cells were collected and subjected to coimmunoprecipitation assays. L, Docking model of SAICAR and PPIMA (left) and interactions of SAICAR with PPM1A (right) in its pocket. Molecular forces implied in the interactions are hydrogen bonds with Arg33, Asp66, and Ala192 and ionic bonds with His62, Lys165, and Arg186. M, The binding of SAICAR to purified PPM1A was analyzed by SPR. The apparent dissociation constant K d is 244 μmol/L. N and O, PPM1A-KD naïve CD4 + T cells were created by lentivirus. Statistics showing the mRNA expression of Foxp3 ( n = 3; N ) and the percentage of Treg cells in CD4 + T cells ( n = 3; O ) by qPCR and flow cytometry after presence in the Treg differentiation condition. Error bars, means ± SD. Statistical significance was assessed using the Student two-tailed t test, one-way ANOVA, and Tukey HSD test. FDR was used in A . *, P < 0.05; **, P < 0.01; ****, P < 0.0001; ns, not significant.

Article Snippet: Material used for the experiment was obtained from the following manufacturers: SAICAR (MedChemExpress), PPM1A (MedChemExpress), ultrapure water, 10 mmol/L Glycine-HCl (pH 2), IPA (MedChemExpress), NHS (Thermo Fisher Scientific), EDC (Thermo Fisher Scientific), and Sensor Chip COOH (Nicoya).

Techniques: Binding Assay, Gene Expression, RNA Sequencing, Cell Culture, Expressing, Negative Control, Isolation, Purification, Flow Cytometry, Two Tailed Test

Journal: Antioxidants

Article Title: Regioselective and Stereoselective Epoxidation of n-3 and n-6 Fatty Acids by Fungal Peroxygenases

doi: 10.3390/antiox10121888

Figure Lengend Snippet: Formulae of the n-3 ( 1 – 8 ) and n-6 ( 9 – 11 ) fatty acids used in enzymatic epoxidation reactions: ( 1 ) Hexadeca-7,10,13-trienoic acid (HTA); ( 2 ) Octadeca-6,9,12,15-tetraenoic acid (SDA); ( 3 ) Eicosa-11,14,17-trienoic acid (ETE); ( 4 ) Eicosa-5,8,11,14,17-pentaenoic acid (EPA); ( 5 ) Heneicosa-6,9,12,15,18-pentaenoic acid (HPA); ( 6 ) Docosa-7,10,13,16,19-pentaenoic acid (DPA); ( 7 ) Docosa-4,7,10,13,16,19-hexaenoic acid (DHA); ( 8 ) Tetracosa-6,9,12,15,18,21-hexaenoic acid (nisinic acid); ( 9 ) Eicosa-8,11,14-trienoic acid (DGLA); ( 10 ) Eicosa-5,8,11,14-tetraenoic acid (AA); and ( 11 ) Docosa-7,10,13,16-tetraenoic acid (AdA).

Article Snippet: These compounds −namely hexadeca-7,10,13-trienoic acid (HTA; 1 ), octadeca-6,9,12,15-tetraenoic acid (stearidonic acid, SDA; 2 ), eicosa-11,14,17-trienoic acid (ETE; 3 ), eicosa-5,8,11,14,17-pentaenoic acid (EPA; 4 ), heneicosa-6,9,12,15,18-pentaenoic acid (HPA; 5 ), docosa-7,10,13,16,19-pentaenoic acid (DPA; 6 ), docosa-4,7,10,13,16,19-hexaenoic acid (DHA; 7 ), tetracosa-6,9,12,15,18,21-hexaenoic acid (nisinic acid; 8 ), eicosa-8,11,14-trienoic acid (dihomo-γ-linoleic acid, DGLA; 9 ), eicosa-5,8,11,14-tetraenoic acid (AA; 10 ) and docosa-7,10,13,16-tetraenoic acid (adrenic acid, AdA; 11 )− were supplied by Larodan ( cis isomers in all cases).

Techniques: