Journal: Nature medicine

Article Title: Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture

doi: 10.1038/nm.3585

Figure Lengend Snippet: (a) Schematic of human 11p15.5 CRC amplicon indicating the position of miR-483 within IGF2 (UCSC Genome Browser). (b) Functional analysis of miR-483 , Igf2 or both in Apc -null colon organoids. Tamoxifen-treated Apc flox/flox ; villin-CreER adult colon organoids were infected with appropriate combinations of control LMP retrovirus, Igf2 cDNA-IRES-GFP or lentivirus miR-483/GFP to generate Apc -null (A), Apc -null/ Igf2 (AI), Apc -null/ miR-483 (AM) or Apc -null/ miR-483 / Igf2 (AIM) organoids. Significant high-grade epithelial dysplasia with nuclear pleiomorphism was only observed with miR-483 -overexpressing organoids (AM and AIM) but not A or AI. H&E, day 50 post-infection. Scale bars, 50 μm. (c) Dysplasia index from blinded pathologic analysis reveals marked transformation of miR-483 -containing modules (AM, AIM) versus Apc -null or Apc/Igf2 (AI or AIM vs. A, ** = P < 0.0001 ) versus Igf2 (AI vs. A; AIM vs. AM, * = P < 0.05 ). Each individual data point represents n=4–6 microscopic fields containing viable organoids. Day 50 post-infection. Mean +/− SE. (d) AM but not A cells demonstrated robust in vivo tumorigenicity following s.c. transplantation into NSG mice (day 60 post-implantation, 500,000 cells, passage 5). H&E staining demonstrated poorly differentiated adenocarcinoma, scale bar, 50 μm. Tumors exhibited PCNA-positive high mitotic index and expressed epithelial markers villin and E-cadherin but not the stromal marker smooth-muscle actin (SMA), scale bars, 50 μm. (bottom, right), AM versus Apc -null (“A”) or other two gene combinations, Tumor weight examined at day 60 post s.c. organoid implantation, n=10 tumors/genotype. * = P < 0.0001 . (e) 96-well invasion and proliferation analyses. FACS-sorted EpCAM + disaggregated cells from day 14 organoids of the indicated genotypes were replated at 10,000 cells/well into 96-well air-liquid interface culture to form secondary organoids. After 14 days of additional culture, proliferation and invasion in A, AI, AM and AIM organoids was determined by Calcein Red-Orange, AM fluorescence signal. N=12 wells/genotype, mean+/−SE. For proliferation * = P < 0.00003 and for invasion * = P < 1 × 10 −7 , † = P <0.03 , all vs. the Apc-null “A” module. (f) MiR-483 represses Pdlim2 and Abtb1 RNA in FACS-sorted EpCAM + /GFP + cells from Apc -null colon organoids 10 days after infection with lentivirus encoding miR-483 or negative control miR . N=3 experiments, mean +/− SE, * = P<0.05 . (g) PDLIM2 rescue experiments. AM vs. A organoids were assayed after 14d of culture in 96-well format for invasion and proliferation with either PDLIM2 cDNA overexpression or Pdlim2 shRNA knockdown. * = P < 0.05 . N=12, mean +/− SE. (h) Schematic of mediation of miR-483 -induced dysplasia by Pdlim2 and other potential targets.

Article Snippet: Mouse Igf2 cDNA was obtained from OriGene (Plasmid # MG201606 ).

Techniques: Amplification, Functional Assay, Infection, Control, Transformation Assay, In Vivo, Transplantation Assay, Staining, Marker, Fluorescence, Negative Control, Over Expression, shRNA, Knockdown

Journal: Life Science Alliance

Article Title: Identification of new drugs to counteract anti-spike IgG-induced hyperinflammation in severe COVID-19

doi: 10.26508/lsa.202302106

Figure Lengend Snippet: (A, B, C) Representative data of macrophage activation assay for (A) the PI3Kα inhibitor alpelisib, (B) the PI3Kδ and PI3Kγ inhibitor duvelisib, and (C) the PI3Kδ inhibitor idelalisib.

Article Snippet: Aleplisib (BYL719) , Selleckchem.com , Cat#S1815.

Techniques: Activation Assay

Journal: Life Science Alliance

Article Title: Identification of new drugs to counteract anti-spike IgG-induced hyperinflammation in severe COVID-19

doi: 10.26508/lsa.202302106

Figure Lengend Snippet:

Article Snippet: Aleplisib (BYL719) , Selleckchem.com , Cat#S1815.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software

Journal: Bioconjugate chemistry

Article Title: Synthesis and evaluation of novel Tc-99m labeled probestin conjugates for imaging APN/CD13 expression in vivo.

doi: 10.1021/bc200546b

Figure Lengend Snippet: Figure 2. Radio-HPLC chromatograms of (a) N3S-PEG2-Probestin, (b) ReO-N3S-PEG2-Probestin, (c) 99mTcO-N3S-PEG2-Probestin, and (d) mouse urine collected at 10 min p.i. of 99mTcO-N3S-PEG2- Probestin.

Article Snippet: Fmoc-8amino-3,6-dioxaoctanoic acid (PEG2) was obtained from Peptides International Inc. (Louisville, KY).

Techniques:

Journal: Bioconjugate chemistry

Article Title: Synthesis and evaluation of novel Tc-99m labeled probestin conjugates for imaging APN/CD13 expression in vivo.

doi: 10.1021/bc200546b

Figure Lengend Snippet: Figure 4. Photograph of the HT-1080 tumor-bearing nude mice used for imaging with 99mTcO-N3S-PEG2-Probestin (arrows indicate tumor) and whole body planar images of the same mice at 1 h p.i. ReO-N3S-PEG2-Probestin (1 mg) was coinjected intravenously along with the 99mTcO-N3S- PEG2-Probestin for blocking APN specific uptake of the radioactivity. Planar images were acquired in prone position.

Article Snippet: Fmoc-8amino-3,6-dioxaoctanoic acid (PEG2) was obtained from Peptides International Inc. (Louisville, KY).

Techniques: Imaging, Blocking Assay, Radioactivity

Journal: Bioconjugate chemistry

Article Title: Synthesis and evaluation of novel Tc-99m labeled probestin conjugates for imaging APN/CD13 expression in vivo.

doi: 10.1021/bc200546b

Figure Lengend Snippet: Figure 5. (a) Abdomen planar images of the HT-1080 tumor-bearing nude mice injected with 99mTcO-N3S-PEG2-Probestin at 1 h p.i. ReO-N3S- PEG2-Probestin (1 mg) was coinjected intravenously along with the 99mTcO-N3S-PEG2-Probestin for blocking APN specific uptake of the radioactivity. Planar images were acquired in prone position. (b) Early and (c) delayed abdomen SPECT images of a HT-1080 tumor-bearing nude mouse injected with 99mTcO-N3S-PEG2-Probestin. Labels: K, kidney; L, Liver; I, Intestine; B, Bladder.

Article Snippet: Fmoc-8amino-3,6-dioxaoctanoic acid (PEG2) was obtained from Peptides International Inc. (Louisville, KY).

Techniques: Injection, Blocking Assay, Radioactivity, Single Photon Emission Computed Tomography