Journal: Biomedicines

Article Title: Characterization of Mesothelin Glycosylation in Pancreatic Cancer: Decreased Core Fucosylated Glycoforms in Pancreatic Cancer Patients’ Sera

doi: 10.3390/biomedicines10081942

Figure Lengend Snippet: Lectins used in WB.

Article Snippet: Fluorescein labelled Phaseolus vulgaris Erythroagglutinin (PHA-E) , Galβ4GlcNAcβ2Manα6 (GlcNAcβ4) (GlcNAcβ4Manα3) Manβ4 , 2 µg/mL , FL-1121 (Vector Laboratories).

Techniques: Concentration Assay, Plasmid Preparation

Journal: Stem cells (Dayton, Ohio)

Article Title: Mesenchymal stem cells and endothelial progenitor cells decrease renal injury in experimental swine renal artery stenosis through different mechanisms

doi: 10.1002/stem.1263

Figure Lengend Snippet: Top: Representative images of CM-DiI labeled (red) endothelial progenitor cells (EPC) or mesenchymal stem cells (MSC) in the post-stenotic kidneys of pigs with renal artery stenosis (RAS) 4 weeks after cell delivery. Green shows peanut agglutinin (PA, green arrow), a distal tubular marker, and cyan shows a proximal tubular marker phaseolus vulgaris erythroagglutinin (PHA-E, cyan arrow). EPC showed mainly tubular engraftment (yellow arrow), while MSC tend to integrate into both proximal tubules (yellow arrow) and interstitial area (red arrow). Bottom: Both EPC and MSC improved RBF and GFR in pigs with RAS, yet MSC more effectively restored GFR. *p<0.05 vs. Normal, †p<0.05 vs. RAS. Scale bar=200µm

Article Snippet: Furthermore, frozen kidney sections from pigs infused with cells were stained with the distal tubular marker peanut agglutinin (5ug/ml, Vector) and proximal tubular marker phaseolus vulgaris erythroagglutinin (PHA-E, 5ug/ml, Vector).

Techniques: Labeling, Marker

Journal: Cell reports

Article Title: Mitotic phosphorylation inhibits the Golgi mannosidase MAN1A1

doi: 10.1016/j.celrep.2022.111679

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rhodamine labeled Phaseolus Vulgaris Leucoagglutinin (PHA-L) was purchased from Vector Labs (RL-1112).

Techniques: Recombinant, Labeling, Plasmid Preparation, Knock-Out, Software

Journal: medRxiv

Article Title: mRNA aggregates harness danger response for potent cancer immunotherapy

doi: 10.1101/2023.03.12.23287108

Figure Lengend Snippet: a , Cryo-electron micrographs. b , nanosight size distributions. c , zeta potential measurements. d , encapsulation efficiency of RNA-LPA at 1:15 mass ratios. e , RNA-LPA cluster growth over time at increasing LP:RNA ratios. f , IFN-α measurements from serum of Balb/c mice (n= 3) within 6h of i.v. RNA-LPA loaded with total tumor derived mRNA from K7M2 (upper limit of detection for assay is 2,000 pg/mL). g , Activated DCs. h , central memory CD4 T cells. i , central memory CD8 T cells from spleens of Balb/c mice (n= 3-5) bearing K7M2 pulmonary sarcomas harvested 24h after a third weekly dose of i.v. RNA-LPA loaded with tumor derived mRNA (from K7M2). j , C57Bl/6 mice (n=7-8) were implanted with B16F10-OVA subcutaneously on day 0 and injected with OT-1 cells intravenously next day. Tetramer+ T cells from spleens were harvested 24h after third weekly dose of i.v. OVA specific RNA-LPA. k , Survival curve of Balb/c mice (n= 8) bearing pulmonary K7M2 sarcomas treated with i.v. tumor derived RNA-LPA. l , Survival curve of Balb/c mice (n= 5-8) bearing pulmonary K7M2 sarcomas treated with three weekly i.v. GFP or K7M2 tumor derived RNA-LPA (data is aggregate of two separate experiments). m , Re-challenge of long-term surviving Balb/c mice (n=7-8) previously treated with K7M2 tumor derived RNA-LPA inoculated at 120 days with K7M2 i.v. versus a new cohort of untreated mice. n , Survival curve of SCID Fox-Chase mice on Balb/c background (n= 7-8) bearing pulmonary K7M2 sarcomas treated with three weekly i.v. GFP or K7M2 tumor derived RNA-LPA. o , Survival curve of C57Bl/6 mice (n=14) bearing intracranial KR158b gliomas transduced with luciferase and pp65 and treated i.v. with weekly GFP or pp65 RNA-LPAs. p , Survival curve of C57Bl/6 mice (n= 7-8) implanted neonatally (P1) with K2 midline gliomas expressing H3K27M and treated with H3K27M encoding RNA-LPA. Significance was determined via parametric student’s t-test ( f-j ), and log-rank test ( k-p ). Error bars are reported as the standard error of the mean ( f ) and standard deviation of the mean ( g-j ).

Article Snippet: T cell mitogen Phytohemagglutinin (PHA-P; InvivoGen, cat. inh-phap) was used as the positive control at a final concentration of 1μg/well while CTL-Test Medium alone served as the negative control.

Techniques: Zeta Potential Analyzer, Encapsulation, Derivative Assay, Injection, Transduction, Luciferase, Expressing, Standard Deviation

Journal: medRxiv

Article Title: mRNA aggregates harness danger response for potent cancer immunotherapy

doi: 10.1101/2023.03.12.23287108

Figure Lengend Snippet: a , Cross section of the whole spleen 2 days following i.v. injection with Cre RNA-LPA. Yellow dashed box maps to magnified panels ‘d.’ b , Top row , lower magnification of laminin (FRC marker) and tdtomato signal; bottom rows represent higher magnification of yellow boxed areas. Co-localization between laminin and tdtomato, calculated using voxel-based signal overlap, is displayed as an independent channel in light blue. c . 3D surface-based co-localization between FRCs (laminin+) with tdtomato. Colocalization is displayed as an independent channel in yellow. d , Top row maps to yellow box in panel ‘a’ with 3D overlay of tdtomato with F4/80; bottom row displays two inlaid boxes at higher magnification in 2D. e , Left column shows 2 different tdtomato cells (in 2D) interfacing with macrophages (separate from ‘d’); right column , 3D capture of these cells, rotating to display maximum contact ‘confluence’ points identified via voxel co-localization (blue) f, g , Cytokine/chemokine response panel from C57Bl/6 mice (n=3) treated with i.v. RNA-LPA. h , Absolute counts of peripheral white blood cells from C57Bl/6 mice (n=5) 6h after i.v. RNA-LPA. i , Absolute counts of activated DCs and activated T cells in spleens of C57Bl/6 mice (n=5) harvested 6h after i.v. RNA-LPA. j , RNA sequencing of established KR158b-luc intracranial tumors harvested 24h after a single tumor derived RNA-LPA. k , Survival curve of C57Bl/6 mice (n=7-8) bearing pulmonary K7M2 sarcomas treated i.v. with three weekly RNA-LPAs concomitantly with IFNAR1 mAbs. l , Serum analysis of IFN-α (left), n=3, and survival curve (right) from C57Bl/6 wild-type versus TLR7 knock-out mice (n=5-8) bearing subcutaneous B16F10-OVA melanomas treated i.v. with OVA RNA-LPA. m , Survival curve from C57Bl/6 wild-type versus MYD88 knock-out mice (n= 7-8) bearing pulmonary B16F10-OVA melanomas treated i.v. with OVA RNA-LPA. n , Serum analysis of IFN-α (left), n=3, and survival curve (right) from C57Bl/6 wild-type versus RIG-I knock-out mice (n=7-8) bearing pulmonary B16F10-OVA melanomas treated i.v. with OVA RNA-LPA. Significance determined via parametric student’s t-test ( h, i, l (left), n (left)), Mixed effects models/repeat ANOVA models ( l , right) and log-rank test ( l (right), m, n (right)). Error bars reported as the standard error of the mean ( h, i, l (right)) and the standard deviation of the mean ( l (left), n (left)).

Article Snippet: T cell mitogen Phytohemagglutinin (PHA-P; InvivoGen, cat. inh-phap) was used as the positive control at a final concentration of 1μg/well while CTL-Test Medium alone served as the negative control.

Techniques: Injection, Marker, RNA Sequencing Assay, Derivative Assay, Knock-Out, Standard Deviation

Journal: medRxiv

Article Title: mRNA aggregates harness danger response for potent cancer immunotherapy

doi: 10.1101/2023.03.12.23287108

Figure Lengend Snippet: a , Study schema of MGMT unmethylated glioblastoma patients enrolled to receive RNA-LPA. b, c, d Cytokine/chemokine sera measurements pre, 2, and 6h post-RNA-LPA. e, f, g , Absolute counts of peripheral blood monocytes ( e ) lymphocytes ( f ), and neutrophils ( g ). h, i, j , Fraction of APCs ( h ) and DC subtypes including mDCs ( i ) and pDCs ( j ). k , Mean fluorescent intensity (MFI) of activated CD8 T cells. l , IFN-γ and granzyme B Elispots using unstimulated and pp65 re-stimulated PBMCs from Patient A25. m, n , Flow cytometric analysis of antigen specific T cells in Patients A25 and B42 by tetramer staining for HLA-A2 restricted pp65 epitope. o , Significantly expanded pp65-specific TCRβ in Patient A25 and B42 samples after 2 and 4 RNA-LPA infusions respectively. p , Proposed mechanisms for RNA-LPA mediated innate and adaptive mobilization/activation (Image created with Biorender.com ).

Article Snippet: T cell mitogen Phytohemagglutinin (PHA-P; InvivoGen, cat. inh-phap) was used as the positive control at a final concentration of 1μg/well while CTL-Test Medium alone served as the negative control.

Techniques: Staining, Activation Assay