Journal: Cell reports. Medicine

Article Title: Safety and immunogenicity of SARS-CoV-2 self-amplifying RNA vaccine expressing an anchored RBD: A randomized, observer-blind phase 1 study.

doi: 10.1016/j.xcrm.2023.101134

Figure Lengend Snippet: Figure 3. SARS-CoV-2 IgG and neutralizing antibody responses (A and B) Serum IgG titers against wild-type SARS- CoV-2 RBD protein (A) and pseudovirus neutralizing antibody titers (ID50) against SARS-CoV-2 variants (B) for non-elderly (left side) and elderly (right side) participants (n = 10 per dose group, and n = 6 for the placebo group). Participants received one injection of VLPCOV-01 (0.3, 1.0, or 3.0 mg), 30 mg BNT162b2, or placebo on day 1 (week 0). Logarithmic values are reported as geometric mean titers for serum IgG and neutralizing antibody against pseudovirus. Bars indicate 95% CIs. (C) The correlation between serum neutralizing antibody titers against pseudovirus Wuhan (wild type) and IgG antibody titers against SARS-CoV-2 RBD protein following booster vaccination. Pearson’s product-moment correlation coefficient and p value were calculated following log trans- formation of source data (r = 0.950, p < 0.001).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti-human CD154-FITC (clone: TRAP1) BD Biosciences cat# 555699; RRID: AB_396049 Mouse anti-human CD3-BUV615 (clone: SP34-2) BD Biosciences cat# 751249; RRID: AB_2875266 Mouse anti-human CD4-PE-Cy5.5 (clone: S3.5) Thermo Ficher Scientific cat# MHCD0418; RRID: AB_10376013 Mouse anti-human CD8-BUV563 (clone: RPA-T8) BD Biosciences cat# 612914; RRID: AB_2870200 Mouse anti-human CD27-PE-Cy5 (clone: 1A4CD27) Beckman coulter cat# 6607107 Mouse anti-human CD45RO-BUV805 (clone: UCHL1) BD Biosciences cat# 748367; RRID: AB_2872786 Mouse anti-human IFN-g-BV786 (clone: 4S.B3) BioLegend cat# 502542; RRID: AB_2563882 Mouse anti-human TNF-BV650 (clone: MAb11) BioLegend cat# 502938; RRID: AB_2562741 Rat anti-human IL-13-BV421 (clone: JES10-5A2) BD Biosciences cat# 563580; RRID: AB_2738290 Mouse anti-human IL-21-Ax647 (clone: 3A3-N21) BD Biosciences cat# 560493; RRID: AB_1645421 Mouse anti-human IL-4-PE-Cy7 (clone:8D4-8) BD Biosciences cat# 560672; RRID: AB_1727547 Mouse anti-human IL-17A-BV605 (clone: BL168) BioLegend cat# 512326; RRID: AB_2563887 Rat anti-human IL-2-BUV737 (clone: MQ117H12) BD Biosciences cat# 612836 Mouse anti-human CD107A-BV711 (clone: H4A3) BioLegend cat# 328640; RRID: AB_2565840 Mouse anti-human MIP1b-Alexa700 (clone: D21-1351) BD Biosciences cat# 561278; RRID: AB_10612008 Anti-SARS-CoV-2 Spike RBD Neutralizing Antibody, Human IgG1 (AS35) ACROBiosystems cat# SAD-S35 Anti-SARS-CoV-2 Spike RBD Neutralizing Antibody, Human IgG2 (AS35) ACROBiosystems cat# SAD-S66 Anti-SARS-CoV-2 Spike RBD Neutralizing Antibody, Human IgG3 (AS35) ACROBiosystems cat# SAD-S67 Anti-SARS-CoV-2 Spike RBD Neutralizing Antibody, Human IgG4 (AS35) ACROBiosystems cat# SAD-S68 Mouse anti-Human IgG1 Fc Secondary Antibody, HRP Thermo Fisher Scientific cat# MH1715; RRID: AB_2539710 Mouse Anti-Human IgG2 Fc-BIOT HP6002 SouthernBiotech cat# 9070-08; RRID: AB_2796638 Mouse Anti-Human IgG3 Hinge-BIOT HP6050 SouthernBiotech cat# 9210-08; RRID: AB_2796700 Mouse Anti-Human IgG4 Fc-BIOT HP6025 SouthernBiotech cat# 9200-08; RRID: AB_2796692 Biological samples Human PBMCs This study This study Chemicals, peptides, and recombinant proteins Benzonase Nuclease, Purity >90% MERCK Millipore cat# 70746 (Continued on next page) Cell Reports Medicine 4, 101134, August 15, 2023 e1

Techniques: Injection

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: Current-potential profiles ( a , c ) and calibration curves ( b , d ) of neutralizing antibodies sIgA and IgG were obtained using the GLEIA−based electrochemical immunosensor. Current−potential responses were tested under different concentrations of ( a ) sIgA 100 ng/mL (blue), 20 ng/mL (grey), 5 ng/mL (orange), 1 ng/mL (red), and 0 ng/mL (black) and ( c ) IgG 62.5 U/mL (blue), 25 U/mL (grey), 6.25 U/mL (orange), 2.5 U/mL (red), and 0 U/mL (black). Calibration curves were indicated for sIgA ( b ) and IgG ( d ) as Michaelis–Menten-type functions by a non-linear curve fitted in the graphing software Origin2022 (OriginLab).

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Software

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: Comparisons of concentrations of neutralizing antibody IgG ( a ) and sIgA ( b ), antioxidant activity (indicated by luminescence inhibition rate) ( c ), and protein concentration ( d ) using saliva samples from 10 individuals are presented together. All 10 saliva samples were collected 3 weeks after the second dose of the vaccine.

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Antioxidant Activity Assay, Inhibition, Protein Concentration

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: The four datasets of neutralizing antibodies—IgG and sIgA, antioxidant activity, and protein concentration—measured in 10 samples are depicted individually in a radar chart for each individual. The numbers T1–T10 correspond to the sample numbers 1–10 in . The data are shown as a ratio of the maximum concentration of each measured item.

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Antioxidant Activity Assay, Protein Concentration, Concentration Assay

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: The results of continuous monitoring of the concentration of neutralizing IgG ( B ) and sIgA ( C ) antibodies, antioxidant activity ( D ), and protein concentration ( E ) in saliva and neutralizing IgG concentration in serum ( A ) before and after 1st, 2nd, and 3rd vaccinations in the same individual over time were compared vertically. The abscissa represents the date and time of sampling. 0: at first vaccination; 1a: 4 days later, 1b: 1 week later, 1c: 2 weeks later, and 1d: 3 weeks later; 2a: 4 days after second vaccination; 2b: 1 week later, 2c: 2 weeks later, 2d: 3 weeks later, 2e: 1 month later, 2f: 2 months later, 2g: 3 months later, 2h: 4 months later, 2i: 5 months later, 2j: 6 months later, and 2k: 8 months later; 3a: 3 days after the third vaccination, 3b: 2 weeks later, and 3c: 1 month later. The units on the vertical axis are as follows: ( A , B ) U/mL, ( C ) ng/mL, ( D ) %, and ( E ) mg/mL.

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Concentration Assay, Antioxidant Activity Assay, Protein Concentration, Sampling

Journal: Cell Death & Disease

Article Title: Sphingosine-1-phosphate promotes CD8 T cell exhaustion in breast cancer via exosomal transfer of TGFBR2

doi: 10.1038/s41419-026-08523-5

Figure Lengend Snippet: A Western blot analysis of TGFBR2 along with Alix and CD63 in Whole cell lysate [W], Exosomal fraction [E] and in micro vesicular fraction [M]. B TGFBR2 expression analysis on the surface of exosome using flow cytometry, isolated from S1P induced MCF7. C Exosomal surface ELISA of TGFBR2. Data are mean ± SD, n = 3 biological replicates, ***P ≤ 0.001 (t test). D Schematic representation of exosomal release from cancer cells and transfer of TGFBR2 to the CD8 T Cells surface. E-G CD8 T cell functional cytokine (IL2, Granzyme B, and IFN-γ expression analyzed using ELISA. Data are represented as mean ± SD, n = 3, ns= non-significant, ***P ≤ 0.001 (one-way ANOVA). H T Cell exhaustion marker expression analyzed using flow cytometry in CD8 T Cell incubated with exosomes isolated from S1P treated TGFBR2 silenced Anti PD-L1 blocking antibody treated MCF7. Data are represented as mean ± SD, n = 3, ns= non-significant, ***P ≤ 0.001, ****P ≤ 0.0001 (one-way ANOVA). I CD8 T Cell proliferation measured using CFSE dye dilution assay incubated with exosomes isolated from S1P treated TGFBR2 silenced anti PD-L1 blocking antibody treated MCF7. J TGFBR2 expression analyzed using western blotting in exosomal fraction as well as in whole cell lysate in S1P1, S1P2 and S1P3 silenced S1P treated MCF7. K TGFBR2 on exosomal surface analyzed using Flow cytometry isolated from S1P1, S1P2 and S1P3 silenced S1P treated MCF7. Statistical analysis was performed using GraphPad Prism 8.

Article Snippet: Anti-PD-L1 blocking antibody (71213) was purchased from BPS Bioscience, and anti-TGFBR2 blocking antibody (AF-241-NA) was purchased from R&D Systems.

Techniques: Western Blot, Expressing, Flow Cytometry, Isolation, Enzyme-linked Immunosorbent Assay, Functional Assay, Marker, Incubation, Blocking Assay, Dilution Assay

Journal: bioRxiv

Article Title: Wnts are endothelial cell-derived PKD1/PKD2-dependent autocrine/paracrine vasodilators

doi: 10.64898/2026.03.17.712518

Figure Lengend Snippet: (A) Representative RT-PCR showing that mRNA for PKD1 is present in isolated endothelial cells from tamoxifen-treated Pkd1 fl/fl but absent in isolated endothelial cells from tamoxifen-treated Pkd1 fl/fl :Cdh5(PAC)-CreERT2 mice. Representative of 5 independent experiments for each genotype. (B) Representative Western blots illustrating PKD1, PKD2, AT1, eNOS, Fzd-7 and actin proteins in mesenteric arteries of Pkd1 fl/fl and Pkd1 ecKO mice. (C) Mean data from experiments shown in panel B. Significance was assessed using Student t-tests, n=5 independent mesenteric arterial lysates for each genotype and each protein. (D) Immunofluorescence images (representative of three mesenteric arteries from three mice for each genotype) illustrating that PKD1 (Alexa Fluor 546) is abolished in endothelial cells of en face mesenteric arteries from Pkd1 ecKO mice. CD31 (Alexa Fluor 488) and DAPI are also shown. Scale bars, 50 μm. (E) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular Wnt9b (3 μg/ml) applied in continuous flow in pressurized (80 mmHg) mesenteric arteries from Pkd1 fl/fl and Pkd1 ecKO mice. (F) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular boiled Wnt9b (3 µg/ml) applied under continuous flow (10 dyn/cm 2 ) in pressurized (80 mmHg) mesenteric arteries from Pkd1 fl/fl and Pkd1 ecKO mice. (G) Mean data from experiments shown in panels E and F, and the modulation of flow and Wnt9b-mediated vasodilation by SRI37892 (Fzd-7 Inh, 5 µM). Significance was assessed using a two-way ANOVA with Holm-Sidak post hoc multiple comparisons test. n=10 arteries from 7 mice of each genotype for flow, flow+ Wnt9b and Wnt9b. n=7 arteries from 5 mice for Fzd-7 Inh+flow and Fzd-7 Inh+flow+Wnt9b. n=5 arteries from 5 mice for each genotype for flow + boiled Wnt9b. (H) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular Wnt5a (3 µg/ml). (I) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular boiled Wnt5a (3 µg/ml). (J) Mean data from experiments shown in panels H and I and the modulation of flow and Wnt5a-induced vasodilation by SRI37892 (Fzd-7 Inh, 5 μM). Significance was assessed using two-way ANOVA with Holm-Sidak post hoc multiple comparisons test. n=8 arteries from 6 mice from Pkd1 fl/fl for flow, flow+ Wnt5a. n=5 arteries from 5 mice from Pkd1 ecKO for flow, flow+ Wnt5a. n=5 arteries from 5 mice of each genotype for flow + boiled Wnt5a. n=8 arteries from 6 mice from Pkd1 fl/fl for flow+ Fzd-7 Inh and flow+ Wnt5a+ Fzd-7 Inh.

Article Snippet: Membranes were blocked with 5% milk or 5% BSA and incubated with one of the following primary antibodies: PKD1 (Polycystic Kidney Disease Research Resource Consortium, Baltimore), PKD2 (Alomone), eNOS (Abcam), p-eNOS (Cell Signaling), Wnt9b (R&D Systems), Wnt5a (R&D Systems), AT1 receptor (Alomone) or actin (Cell Signaling) overnight at 4°C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Western Blot, Immunofluorescence

Journal: Neurochemistry international

Article Title: Sexually dimorphic and brain region-specific transporter adaptations in system x c - null mice.

doi: 10.1016/j.neuint.2020.104888

Figure Lengend Snippet: Fig. 1. Comparison of glutamate and cysteine transporter levels between wild-type (þ/þ) and system xc - null (sut/sut) female mice in striatum, hippo campus, and cortex. Plasma membrane striatal, hippocampal, and cortical protein extracted in pairs from +/+ and sut/sut female mice [5 striata or 5 hippocampi (5 mice per 2 samples), or 2 cortices (1 mouse per sample)] were separated under reducing conditions via 8% SDS-PAGE. Representative blots and protein quantification for EAAT1, 2, 3, ASCT1, SNAT3 and LAT2 protein in striatum (A), hippocampus (B) and cortex (C). Protein quantification was performed by normalizing to each samples’ representative total protein to correct for differences in loading followed by baseline correction to the paired +/+ levels. Values are graphed as Tukey’s box and whisker plots with the box representing the 2nd and 3rd quartiles, and whiskers representing points within 1.5 times the interquartile distance. An asterisk (*) represents a significant between-group difference calculated using a ratio-paired t-test on transformed data. Between-group differences for each brain areas are as follows: (A) Striatum: EAAT1 (p = 0.044; n = 6), EAAT2 (p = 0.477; n = 11), EAAT3 (p = 0.234; n = 10), ASCT1 (p = 0.343; n = 14), SNAT3 (p = 0.279; n = 6) and LAT2 (p = 0.376; n = 6). (B) Hippocampus: EAAT1 (p = 0.085; n = 11), EAAT2 (p = 0.169; n = 11), EAAT3 (p = 0.086; n = 7), ASCT1 (p = 0.144; n = 14), SNAT3 (p = 0.213; n = 7) and LAT2 (p = 0.436; n = 7). (C) Cortex: EAAT1 (p = 0.092; n = 14), EAAT2 (p = 0.2391; n = 14), EAAT3 (p = 0.426; n = 14), ASCT1 (p = 0.0267; n = 13), SNAT3 (p = 0.0001; n = 6), and LAT2 (p = 0.260; n = 6).

Article Snippet: Protein Host Species Company Catalog Number RRID Epitope EAAT1 Rabbit Abcam Ab416 AB_304334 C-terminus of rat EAAT1 EAAT2 Rabbit Abcam Ab41621 AB_941789 C-terminus of rat EAAT2 EAAT3 Rabbit Abcam Ab124802 AB_10974334 Human EAAT3 residues 150-250 ASCT1 Rabbit Alomone Labs ANT-081 AB_2756719 Mouse Slc1a4 residues 500-512 LAT2 Mouse OriGene TA500514 AB_11124222 Full length Slc7a8 from HEK293T cells SNAT3 Rabbit Proteintech 14315-1-AP AB_941782 Residues 1–191 of Slc38a3 fusion protein SOD1 Rabbit Millipore 07–043 AB_310587 C-terminus of human Cu/Zn SOD SOD2 Rabbit Sigma Aldrich HPA001814 AB_1080134 Residues 2–146 of human SOD2 Catalase Mouse Sigma Aldrich C0979 AB_258720 Human erythrocyte catalase H.M. Sosnoski et al. Neurochemistry International 141 (2020) 104888 comparisons that controlled for procedural variabilities in the amount of protein extracted.

Techniques: Comparison, Clinical Proteomics, Membrane, SDS Page, Whisker Assay, Transformation Assay

Journal: Neurochemistry international

Article Title: Sexually dimorphic and brain region-specific transporter adaptations in system x c - null mice.

doi: 10.1016/j.neuint.2020.104888

Figure Lengend Snippet: Fig. 2. Comparison of glutamate and cysteine transporter levels between wild-type (þ/þ) and system xc - null (sut/sut) male mice in striatum, hippo campus, and cortex. Plasma membrane striatal, hippocampal, and cortical protein extracted in pairs from +/+ and sut/sut male mice [5 striata or 5 hippocampi (5 mice per 2 samples), or 2 cortices (1 mouse per sample)] were separated under reducing conditions via 8% SDS-PAGE. Representative blot for detection of EAAT1, 2, 3, ASCT1, SNAT3 and LAT2 protein in striatum (A), hippocampus (B), and cortex (C). Protein quantification was performed by normalizing to each samples’ representative total protein levels to correct for differences in loading followed by baseline correction to the paired +/+ levels. Values are graphed as Tukey’s box and whisker plots with the box representing the 2nd and 3rd quartiles, and whiskers representing points within 1.5 times the interquartile distance. An asterisk (*) represents a significant between-group difference calculated using a ratio-paired t-test on transformed data. (A) Striatum: EAAT1 (p = 0.3866; n = 7), EAAT2 (p = 0.468; n = 7), EAAT3 (p = 0.351; n = 6), ASCT1 (p = 0.493; n = 7), SNAT3 (p = 0.0267; n = 7) and LAT2 (p = 0.465; n = 7). (B) Hippocampus: EAAT1 (p = 0.186; n = 7), EAAT2 (p = 0.079; n = 7), EAAT3 (p = 0.487; n = 6), ASCT1 (p = 0.469; n = 7), SNAT3 (p = 0.438; n = 7) and LAT2 (p = 0.386; n = 6). (C) Cortex: EAAT1 (p = 0.149; n = 7), EAAT2 (p = 0.161; n = 7), EAAT3 (p = 0.325; n = 7), ASCT1 (p = 0.429; n = 7), SNAT3 (p = 0.026; n = 5) and LAT2 (p = 0.254; n = 5).

Article Snippet: Protein Host Species Company Catalog Number RRID Epitope EAAT1 Rabbit Abcam Ab416 AB_304334 C-terminus of rat EAAT1 EAAT2 Rabbit Abcam Ab41621 AB_941789 C-terminus of rat EAAT2 EAAT3 Rabbit Abcam Ab124802 AB_10974334 Human EAAT3 residues 150-250 ASCT1 Rabbit Alomone Labs ANT-081 AB_2756719 Mouse Slc1a4 residues 500-512 LAT2 Mouse OriGene TA500514 AB_11124222 Full length Slc7a8 from HEK293T cells SNAT3 Rabbit Proteintech 14315-1-AP AB_941782 Residues 1–191 of Slc38a3 fusion protein SOD1 Rabbit Millipore 07–043 AB_310587 C-terminus of human Cu/Zn SOD SOD2 Rabbit Sigma Aldrich HPA001814 AB_1080134 Residues 2–146 of human SOD2 Catalase Mouse Sigma Aldrich C0979 AB_258720 Human erythrocyte catalase H.M. Sosnoski et al. Neurochemistry International 141 (2020) 104888 comparisons that controlled for procedural variabilities in the amount of protein extracted.

Techniques: Comparison, Clinical Proteomics, Membrane, SDS Page, Whisker Assay, Transformation Assay