Journal: Nutrients

Article Title: Integrated Proteomics and Metabolomics Reveal the Direct Hepatic Protection of Propionate Against Alcoholic Liver Disease via the RGN-PPARα Pathway

doi: 10.3390/nu18050872

Figure Lengend Snippet: The effect of PA on lipid accumulation in ALD mice. ( A ) Schematic of the acute alcoholic liver injury model in mice with propionate treatment. ( B , C ) Serum ALT and AST levels in mice. ( D , E ) H&E and Oil Red O staining of liver tissues (scale bar: 50 μm). ( F ) The relative area of Oil Red O staining. ( G , H ) Liver levels of TC and TG in mice. ( I – L ) Serum concentrations of TC, TG, HDL-C and LDL-C in mice. ( M ) Representative Western blot analysis and ( N ) quantification of ACC1, SREBP1 and CPT1A protein expression in liver tissues normalized to β-actin (n = 4). The data are presented as means ± SEM. * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies, including β-actin (#66009-1-Ig), CPT1A (#15184-1-AP), SREBP-1 (#14088-1-AP), ACC1 (#21923-1-AP), ACOX1 (#83731-2-RR), PPARα (#66826-1-Ig) and RGN (#17947-1-AP), were obtained from Proteintech Group (Wuhan, China).

Techniques: Staining, Western Blot, Expressing

Journal: Nutrients

Article Title: Integrated Proteomics and Metabolomics Reveal the Direct Hepatic Protection of Propionate Against Alcoholic Liver Disease via the RGN-PPARα Pathway

doi: 10.3390/nu18050872

Figure Lengend Snippet: Propionate suppressed cell injury and lipid accumulation in vitro. ( A ) Changes in AML-12 cell viability after a series of doses of ethanol challenge. ( B ) Changes in AML-12 cell viability after a series of concentrations of OA incubation. ( C ) Effects of different concentrations of propionate on the viability of AML-12 cells. ( D ) Propionate restored AML-12 cells’ viability following co-treatment with 200 mM ethanol and 0.5 mmol/L OA. ( E ) Representative views of Oil Red O staining of AML-12 cells (scale bars: 100 μm and 50 μm). ( F ) Representative images of Western blot analysis and ( G ) quantification of ACC1, SREBP1 and CPT1A protein expression in AML-12 cells normalized to β-actin (n = 3). The data are presented as means ± SEM. * p < 0.05; ** p < 0.01.

Article Snippet: Primary antibodies, including β-actin (#66009-1-Ig), CPT1A (#15184-1-AP), SREBP-1 (#14088-1-AP), ACC1 (#21923-1-AP), ACOX1 (#83731-2-RR), PPARα (#66826-1-Ig) and RGN (#17947-1-AP), were obtained from Proteintech Group (Wuhan, China).

Techniques: In Vitro, Incubation, Staining, Western Blot, Expressing

Journal: The Journal of investigative dermatology

Article Title: Expression Differences in BCL2 Family Members between Uveal and Cutaneous Melanomas Account for Varying Sensitivity to BH3 Mimetics.

doi: 10.1016/j.jid.2021.11.035

Figure Lengend Snippet: Figure 2. BFL1 contributes to UM’s sensitivity to MCL1i. (a, b) Immunoblot showing the endogenous expression of (a) antiapoptotic and (b) BH3-only proapoptotic BCL2 family of proteins in a panel of CM and UM cell lines. (c) Quantification of the immunoblot data of a and b. (d) ATP assay data of single-drug treatments of S63845 on control and BFL1 OE UM cell lines. Y-axis shows the percentage of relative viability, and the X-axis indicates the dosages of the drug in micrometer. (e) Immunoblots confirming the overexpression. Error bars represent SEM for all figures. * indicates P < 0.05; ** indicates P < 0.01. CM, cutaneous melanoma; MCL1i, MCL1 inhibitor; OE, overexpressed; UM, uveal melanoma.

Article Snippet: Creation of BFL1-overexpressed cell lines UM cells were transduced with BFL1 (NM_004049) Human Tagged ORF Clone Lentiviral Particles (#RC201965L4V) or Lenti-ORF Con- trol Particles (#PS100093V) (Origene Technologies, Rockville, MD) per the manufacturer’s instructions.

Techniques: Western Blot, Expressing, ATP Assay, Control, Over Expression

Journal: The Journal of investigative dermatology

Article Title: Expression Differences in BCL2 Family Members between Uveal and Cutaneous Melanomas Account for Varying Sensitivity to BH3 Mimetics.

doi: 10.1016/j.jid.2021.11.035

Figure Lengend Snippet: Figure 4. TCGA data indicate the clinical relevancy of differential expression of BCL2 proteins between CM and UM and lower BFL1 and higher PUMA in UM than in other cancers. (a) The expression data for all the eight genes analyzed to compare the expressions between UM and CM. Of all eight genes listed, the first seven genes are significantly different between the two subtypes. Expression data of (b) BFL1 and (c) PUMA across various cancers are shown. For b and c, the mRNA level was compared for UM with those for each of the other types of cancers. *** indicates the least significant P-value of the comparison at P < 0.001. For b, pheochromocytoma, diffuse glioma, and prostate adenocarcinoma are additionally significant (P < 0.01) compared with UM. CM, cutaneous melanoma; TCGA, The Cancer Genome Atlas; UM, uveal melanoma.

Article Snippet: Creation of BFL1-overexpressed cell lines UM cells were transduced with BFL1 (NM_004049) Human Tagged ORF Clone Lentiviral Particles (#RC201965L4V) or Lenti-ORF Con- trol Particles (#PS100093V) (Origene Technologies, Rockville, MD) per the manufacturer’s instructions.

Techniques: Quantitative Proteomics, Expressing, Comparison

Journal: PLoS ONE

Article Title: Intrinsic Resistance to 5-Fluorouracil in a Brain Metastatic Variant of Human Breast Cancer Cell Line, MDA-MB-231BR

doi: 10.1371/journal.pone.0164250

Figure Lengend Snippet: (A) Gene expression levels in MDA-MB-231 (Parent), MDA-MB-231SCP2 (SCP2), and MDA-MB-231BR (BR). Candidate genes for drug resistance were screened by RNA sequencing. The intensity of gene expression in the heat map is shown relative to that in the Parent. (B) COX-2 and BCL2A1 mRNA and protein expression levels in Parent, SCP2, and BR. mRNA expression levels in the SCP2 and BR are shown relative to those in the Parent. Data represent the mean with SEM of at least three independent samples (*** P <0.001 vs . Parent and SCP2) for mRNA expressions.

Article Snippet: pCMV-Myc-DDK-Entry vector or pCMV-Myc-DDK- BCL2A1 vector (Origene Technologies, Inc., Rockville, MD) was transfected into MDA-MB-231 and T-47D using Lipofectamine ® LTX, PLUS ™ reagent, and Opti-MEM reduced serum media, according to the manufacturer's protocol (ThermoFisher Scientific Inc.).

Techniques: Expressing, RNA Sequencing Assay

Journal: PLoS ONE

Article Title: Intrinsic Resistance to 5-Fluorouracil in a Brain Metastatic Variant of Human Breast Cancer Cell Line, MDA-MB-231BR

doi: 10.1371/journal.pone.0164250

Figure Lengend Snippet: (A) Cytotoxicity of 5-FU to MDA-MB-231BR in the presence of 1 μM BAY11-7082. Blue broken lines parallel to x-axis represent the cell viability of MDA-MB-231 (Parent) treated with 30 μM 5-FU. Data represent the mean with SEM of four independent samples. (B) Changes in protein expressions of COX-2 and BCL2A1 in MDA-MB-231BR in response to 1 μM BAY11-7082.

Article Snippet: pCMV-Myc-DDK-Entry vector or pCMV-Myc-DDK- BCL2A1 vector (Origene Technologies, Inc., Rockville, MD) was transfected into MDA-MB-231 and T-47D using Lipofectamine ® LTX, PLUS ™ reagent, and Opti-MEM reduced serum media, according to the manufacturer's protocol (ThermoFisher Scientific Inc.).

Techniques:

Journal: PLoS ONE

Article Title: Intrinsic Resistance to 5-Fluorouracil in a Brain Metastatic Variant of Human Breast Cancer Cell Line, MDA-MB-231BR

doi: 10.1371/journal.pone.0164250

Figure Lengend Snippet: (A) Cytotoxicity of 5-FU to MDA-MB-231BR after transient transfection with BCL2A1 siRNA. Blue broken lines parallel to x-axis represent the cell viability of MDA-MB-231 (Parent) treated with 30 μM 5-FU. Data represent the mean with SEM of three independent samples (* P <0.05, BR treated with 30 μM 5-FU and siControl vs . BR treated with 30 μM 5-FU and si BCL2A1 ). (B) Changes in protein expression of BCL2A1 in MDA-MB-231BR in response to BCL2A1 siRNA.

Article Snippet: pCMV-Myc-DDK-Entry vector or pCMV-Myc-DDK- BCL2A1 vector (Origene Technologies, Inc., Rockville, MD) was transfected into MDA-MB-231 and T-47D using Lipofectamine ® LTX, PLUS ™ reagent, and Opti-MEM reduced serum media, according to the manufacturer's protocol (ThermoFisher Scientific Inc.).

Techniques: Transfection, Expressing

Journal: PLoS ONE

Article Title: Intrinsic Resistance to 5-Fluorouracil in a Brain Metastatic Variant of Human Breast Cancer Cell Line, MDA-MB-231BR

doi: 10.1371/journal.pone.0164250

Figure Lengend Snippet: BCL2A1 protein expression of (A) MDA-MB-231 and (C) T-47D after transfection with a pCMV-Myc-DDK Entry vector (Vector) or a pCMV-Myc-DDK BCL2A1 vector ( BCL2A1 ). Cytotoxicity of 5-FU toward (B) MDA-MB-231 transfected with a BCL2A1 vector and (D) T-47D transfected with a BCL2A1 vector. The insets show the IC 50 of 5-FU against cell lines transfected with vectors. Data represent the mean with SEM of at least three independent samples (*** p <0.001 vs . MDA-MB-231 transfected with a control vector; * p <0.05 vs . T-47D transfected with a control vector).

Article Snippet: pCMV-Myc-DDK-Entry vector or pCMV-Myc-DDK- BCL2A1 vector (Origene Technologies, Inc., Rockville, MD) was transfected into MDA-MB-231 and T-47D using Lipofectamine ® LTX, PLUS ™ reagent, and Opti-MEM reduced serum media, according to the manufacturer's protocol (ThermoFisher Scientific Inc.).

Techniques: Expressing, Transfection, Plasmid Preparation

Journal: PLoS ONE

Article Title: Intrinsic Resistance to 5-Fluorouracil in a Brain Metastatic Variant of Human Breast Cancer Cell Line, MDA-MB-231BR

doi: 10.1371/journal.pone.0164250

Figure Lengend Snippet: Protein expression of BCL2A1 in various human cell lines.

Article Snippet: pCMV-Myc-DDK-Entry vector or pCMV-Myc-DDK- BCL2A1 vector (Origene Technologies, Inc., Rockville, MD) was transfected into MDA-MB-231 and T-47D using Lipofectamine ® LTX, PLUS ™ reagent, and Opti-MEM reduced serum media, according to the manufacturer's protocol (ThermoFisher Scientific Inc.).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Intrinsic Resistance to 5-Fluorouracil in a Brain Metastatic Variant of Human Breast Cancer Cell Line, MDA-MB-231BR

doi: 10.1371/journal.pone.0164250

Figure Lengend Snippet: Celecoxib acts on COX-2 but did not regulate BCL2A1 expression, while BAY11-7082 acts directly on NF-κB and moderately regulated BCL2A1 expression, and neither of these sensitized MDA-MB-231BR to 5-FU. Both ABT-263 and BCL2A1 siRNA directly regulate BCL2A1, and were effective for the sensitization of MDA-MB-231BR to 5-FU.

Article Snippet: pCMV-Myc-DDK-Entry vector or pCMV-Myc-DDK- BCL2A1 vector (Origene Technologies, Inc., Rockville, MD) was transfected into MDA-MB-231 and T-47D using Lipofectamine ® LTX, PLUS ™ reagent, and Opti-MEM reduced serum media, according to the manufacturer's protocol (ThermoFisher Scientific Inc.).

Techniques: Expressing

Journal: Nature communications

Article Title: Distinctive phenotypes and functions of innate lymphoid cells in human decidua during early pregnancy.

doi: 10.1038/s41467-019-14123-z

Figure Lengend Snippet: Fig. 6 dNK responses triggered by activating KIR. a Representative XCL1 and CD107a staining by CyTOF in lineage negative (Lin−) CD56+ decidual cells co-expressing 1–3 Killer-cell immunoglobulin-like receptors (KIR) following a 6 h co-culture with K562 (n = 10). b Representative XCL1 and CD107a staining by mass cytometry in Lin-CD56+ cells co-expressing 1–3 KIR following 4 h stimulation by PMA plus ionomycin (n = 8). c Representative XCL1 staining by flow cytometry in Lin-CD56+KIR2DS4+ decidual cells co-expressing 1–3 additional KIRs following activation via P815 cells coated with anti-KIR2DS4 (n = 6). d Correlation of frequency of XCL1+Lin-CD56+ decidual cells and mean side scatter for the same subset (n = 20). Two-tailed p-value calculated for Pearson correlation coefficients. Two-tailed one-way ANOVA of matched data points with Tukey correction and 95% confidence level was used. *p < 0.05, **p < 0.01, ***p < 0.001. Source data are provided as a Source Data file.

Article Snippet: The murine cell line P815 and human cell line K562 were purchased from DSMZ in Germany.

Techniques: Staining, Expressing, Co-Culture Assay, Mass Cytometry, Cytometry, Activation Assay, Two Tailed Test

Journal: Annals of Hematology

Article Title: The BET bromodomain inhibitor ZEN-3365 targets the Hedgehog signaling pathway in acute myeloid leukemia

doi: 10.1007/s00277-021-04602-z

Figure Lengend Snippet: Combinatorial treatment of AML reporter cell lines with ZEN-3365 and GANT-61 decreases GLI reporter activity in vitro. a MV4-11, b MOLM13, c Kasumi-1, and d HL60 reporter cell lines were treated with 2500 or 5000 nM GANT-61 and 50 nM or 100 nM ZEN-3365 for 24 h. Significance calculated against GANT-61 single treatment, * p < 0.05, ** p < 0.01, and ns = not significant

Article Snippet: The following human AML cell lines were used: MV4-11 (ATCC, #CRL9591, RRID:CVCL_0064), MOLM13 (DSMZ, #ACC554, RRID:CVCL_2119), HL60 (DSMZ, #ACC3, RRID:CVCL_002), KG-1 (DSMZ, #ACC14, RRID:CVCL_0374), Kasumi-1 (DSMZ, #ACC220, RRID_CVCL_0589), OCI-AML3 (DSMZ, #ACC582, RRID:CVCL_1844).

Techniques: Activity Assay, In Vitro

Journal: Annals of Hematology

Article Title: The BET bromodomain inhibitor ZEN-3365 targets the Hedgehog signaling pathway in acute myeloid leukemia

doi: 10.1007/s00277-021-04602-z

Figure Lengend Snippet: ZEN-3365 and GANT-61 present antiproliferative effects. GANT-61 (5 or 10 μM) and ZEN-3365 (100 or 200 nM) inhibit cell proliferation over 7 days for a MV4-11, b HL60, c KG-1, d MOLM13, e Kasumi-1, and f OCI-AML3. g Combinatorial treatment of primary AML sample with GANT-61 (5 or 10 M) and ZEN-3365 (100 or 200 nM) leads to a decreased cell proliferation after 4 days. Significance calculated against GANT-61 treatment alone, * p <0.05, ** p <0.01, and ns=not significant

Article Snippet: The following human AML cell lines were used: MV4-11 (ATCC, #CRL9591, RRID:CVCL_0064), MOLM13 (DSMZ, #ACC554, RRID:CVCL_2119), HL60 (DSMZ, #ACC3, RRID:CVCL_002), KG-1 (DSMZ, #ACC14, RRID:CVCL_0374), Kasumi-1 (DSMZ, #ACC220, RRID_CVCL_0589), OCI-AML3 (DSMZ, #ACC582, RRID:CVCL_1844).

Techniques:

Journal: Annals of Hematology

Article Title: The BET bromodomain inhibitor ZEN-3365 targets the Hedgehog signaling pathway in acute myeloid leukemia

doi: 10.1007/s00277-021-04602-z

Figure Lengend Snippet: The colony formation potential of AML cell lines is decreased under single or combinatorial treatment. a MV4-11, b MOLM13, c OCI-AML3, and d HL60 were treated with 5 μM GANT-61 and 0.1 μM ZEN-3365 or in combination and cultured in methocult medium for one week. Colony numbers were counted with an inverted microscope. * p <0.05, ** p <0.01, and ns=not significant

Article Snippet: The following human AML cell lines were used: MV4-11 (ATCC, #CRL9591, RRID:CVCL_0064), MOLM13 (DSMZ, #ACC554, RRID:CVCL_2119), HL60 (DSMZ, #ACC3, RRID:CVCL_002), KG-1 (DSMZ, #ACC14, RRID:CVCL_0374), Kasumi-1 (DSMZ, #ACC220, RRID_CVCL_0589), OCI-AML3 (DSMZ, #ACC582, RRID:CVCL_1844).

Techniques: Cell Culture, Inverted Microscopy