Journal: PLoS ONE

Article Title: Combined Bezafibrate and Medroxyprogesterone Acetate: Potential Novel Therapy for Acute Myeloid Leukaemia

doi: 10.1371/journal.pone.0008147

Figure Lengend Snippet: ( A ) HL60 cells were treated for 7 days with solvent control, B/M alone and B/M combined with either 1 nM ATRA, 1 nM 1α,25(OH) 2 vitamin D 3 (D 3 ) or both. A representative histogram of CD11b flow cytometry is shown in the left panel and results from N = 4 experiments in the middle panel . Mean is indicated by the black bar. ( B ) Cell morphology was analysed by Jenner-Giemsa staining of cytospins. Differentiated neutrophils are identified by classical poly-lobed nuclei (arrows) and apoptotic cells highlighted by asterix. Statistics: *p<0.001. ( C ) Primary AML cells taken from a karyotypically normal non-APL AML were treated with solvent CONT, B/M±1nmATRA/1nM D3. Differentiation was determined by CD11b staining and flow cytometry and by morphological analysis of Jenner-Giemsa stained cytospins after 18 days of treatment. ( D ) ROS generation was measured as described above in primary AML cells treated with cont, B/M±1nmATRA/1nM D3 for 48 hours.

Article Snippet: Bezafibrate (0.5 M in DMSO), MPA (5 mM in ethanol), ATRA (1 mM in DMSO), and 1α, 25(OH) 2 vitamin D 3 (1 mM in ethanol) (Sigma Aldrich).

Techniques: Flow Cytometry, Staining

Journal: Nature Cancer

Article Title: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia

doi: 10.1038/s43018-023-00622-y

Figure Lengend Snippet: a , b , RT-qPCR analysis of Ucp1 ( a ) and beiging markers ( b ) in SVF-differentiated adipocytes treated with 0 nM (DMSO only), 10 nM, 100 nM or 1 µM ATRA for 24 hours (n = 3 biologically independent experiments). c , RT-qPCR analysis of Arg1 in BMDMs treated with ATRA for 72 hours (n = 3 biologically independent experiments). d , RT-qPCR analysis of Arg1 and Mrc1 in BMDMs treated with recombinant IL-33 for 72 hours (n = 3 biologically independent experiments). Data shown represent mean ± SEM, 1-way ANOVA (a), 2-way ANOVA (b) or unpaired two-sided t-test with Welch correction (c,d).

Article Snippet: After 7 d, cells were serum starved overnight and treated with recombinant mouse IL-33 (250 ng ml –1 ; Peprotech) or ATRA (1 µM; Cayman Chemical) for 72 h.

Techniques: Quantitative RT-PCR, Recombinant

Journal: Nature Cancer

Article Title: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia

doi: 10.1038/s43018-023-00622-y

Figure Lengend Snippet: a , RT–qPCR analysis of IGFBP3 mRNA levels in AdN1ICD-overexpressing human AT-ECs compared to AdGFP controls ( n = 5 (vWAT ECs) or 9 (vWAT ECs) biologically independent experiments). b , c , Representative western blot ( b ) and quantification ( c ) of IGFBP3 protein levels normalized to VCP ( n = 8 (vWAT ECs) or 12 (sWAT ECs) biologically independent experiments). d , Igfbp3 mRNA levels in NICD iOE-EC AT-ECs isolated from male mice at 2 weeks after tamoxifen treatment ( n = 6 animals per group). e , f , RT–qPCR ( e ) and western blotting ( f ) of IGFBP3 in human AT-ECs treated with 0 nM (DMSO only), 10 nM, 100 nM or 1 µM ATRA. The western blot image is representative of three individual experiments; n = 3 biologically independent experiments. g , IGFBP3 protein levels were quantified relative to VCP ( n = 3 biologically independent experiments). h , RT–qPCR analysis of IGFBP3 expression in human vWAT and sWAT ECs after treatment with 1, 2.5 or 5 µM RAR antagonist BMS195614 or DMSO ( n = 3 biologically independent experiments). i , j , RT–qPCR analysis of Igfbp3 levels in isolated NICD iOE-EC vWAT and sWAT adipocytes ( n = 3–5 animals per group; i ) as well as whole sWAT ( j ) at 4 and 7 weeks after recombination ( n = 3 (week 4) or 4 (week 7) animals per group). k , IGFBP3 (DAB) immunohistochemical stainings of sWAT from NICD iOE-EC mice at 7 weeks after recombination; scale bar, 100 µm. Images are representative of two individual experiments. l , IGFBP3 + nuclei were quantified as a percentage of total nuclei ( n = 7–9 animals per group pooled from two independent cohorts). m , n , Western blot ( m ) and quantification ( n ) of cleaved caspase-3 levels of SVF-differentiated adipocytes treated with recombinant IGFBP3 (100 ng ml –1 ) for 72 h. Data were normalized to VCP ( n = 3 biologically independent experiments). o , Apoptosis (PS) and necrosis (7-AAD) were assessed using the Apoptosis/Necrosis Assay kit from Abcam ( n = 4 biologically independent experiments; shown are biological replicates representing the averages of five technical replicates). Data shown represent mean ± s.e.m. and were analyzed by unpaired, two-sided t -test with Welch correction ( a , c , d , i and j ), two-way analysis of variance (ANOVA) with Dunnett’s test ( e , g and h ), Mann–Whitney test ( l and n ) or Sidak’s multiple comparisons test ( o ). The experiment in d was performed twice with consistent results. Shown is one representative experiment. Experiments in k and l were performed in two independent cohorts, and results were pooled. Results were consistent between the two experiments.

Article Snippet: After 7 d, cells were serum starved overnight and treated with recombinant mouse IL-33 (250 ng ml –1 ; Peprotech) or ATRA (1 µM; Cayman Chemical) for 72 h.

Techniques: Quantitative RT-PCR, Western Blot, Isolation, Expressing, Immunohistochemistry, Recombinant, MANN-WHITNEY

Journal: Frontiers in Microbiology

Article Title: Susceptibility of Pseudomonas aeruginosa Biofilm to Alpha-Helical Peptides: D-enantiomer of LL-37

doi: 10.3389/fmicb.2011.00128

Figure Lengend Snippet: Peptides used in this study .

Article Snippet: The anti-microbial activity of the NA-CATH and NA-CATH:ATRA1-ATRA1 (AAPPTEC, Louisville, KY, USA), the variations on the ATRA peptides (Genscript, Piscataway, NJ, USA), LL-37 (AnaSpec 61302), and D-LL-37 (Lifetein, South Plainfield, NJ, USA) against P. aeruginosa (ATCC 19429) were determined as previously described, with some modification (Han et al., ; Papanastasiou et al., ).

Techniques: Sequencing

Journal: Frontiers in Microbiology

Article Title: Susceptibility of Pseudomonas aeruginosa Biofilm to Alpha-Helical Peptides: D-enantiomer of LL-37

doi: 10.3389/fmicb.2011.00128

Figure Lengend Snippet: EC50s of AMPs against P. aeruginosa .

Article Snippet: The anti-microbial activity of the NA-CATH and NA-CATH:ATRA1-ATRA1 (AAPPTEC, Louisville, KY, USA), the variations on the ATRA peptides (Genscript, Piscataway, NJ, USA), LL-37 (AnaSpec 61302), and D-LL-37 (Lifetein, South Plainfield, NJ, USA) against P. aeruginosa (ATCC 19429) were determined as previously described, with some modification (Han et al., ; Papanastasiou et al., ).

Techniques: Molecular Weight

Journal: Viruses

Article Title: All-Trans Retinoic Acid Exhibits Antiviral Effect against SARS-CoV-2 by Inhibiting 3CLpro Activity

doi: 10.3390/v13081669

Figure Lengend Snippet: ATRA inhibits SARS-CoV-2 replication ( A ) Assessing the antiviral effect of ATRA in VeroE6/TMPRSS2 cells. VeroE6/TMPRSS2 cells treated with ATRA (0.1–100 µM) for 3 h prior to infection were infected with SARS-CoV-2 (MOI = 0.05) and washed off the unbound virus at 2 h of infection. Cell viability assay and RT-PCR were performed on the attached cells and supernatant culture medium respectively, 48 h after infection. ( B ) The results of the cell viability assay. IC 50 was calculated for the inhibition of SARS-CoV-2 induced cell death. Error bars obtained from triplicate testing. p -value < 0.005 (***) ( C ) Viral quantification by RT-PCR. Error bars obtained from duplicate testing. p -value < 0.005 (***) ( D ) Immunostaining of SARS-CoV-2 N protein in infected cells. VeroE6/TMPRSS2 cells treated with ATRA (1, 10, 25 µM) 3 h before infection were infected with SARS-CoV-2 (MOI = 0.05). After 24 h of infection, cells were fixed with 4% paraformaldehyde and immunostained. Red stained cells represent SARS-CoV-2 N protein, blue-stained nuclei with DAPI. ( E ) Schematic representation of the time of addition assay of ATRA. ( F - H ) The results of RT-PCR for ATRA ( F ), Remdesivir ( G ) and Camostat ( H ). The result of full-time colored grey, entry colored blue, and post-entry colored red. Error bars obtained from duplicate testing. p -value < 0.005 (***).

Article Snippet: We compared the efficacy of ATRA (1, 10, 25 µM) against 10 µM of remdesivir [ ].

Techniques: Infection, Viability Assay, Reverse Transcription Polymerase Chain Reaction, Inhibition, Immunostaining, Staining