Journal: Science (New York, N.Y.)

Article Title: A tool kit of highly selective and sensitive genetically encoded neuropeptide sensors

doi: 10.1126/science.abq8173

Figure Lengend Snippet: (A) Primary cultured rat cortical neurons expressing SST1.0 (A1) or CRF1.0 (A2), showing sensor expression (left), pseudocolor responses (middle), and cell membrane localization (right). Scale bars, 100 μm (left) and 20 μm (right). (B) Example fluorescence traces (left) and summary data (right) of neurons expressing SST1.0 (B1) or CRF1.0 (B2); where indicated, peptides and antagonists were applied (n = 66 to 115 ROIs from three or four coverslips). (C) Normalized dose-response curves of neurons expressing SST1.0 (C1) or CRF1.0 (C2) in response to the indicated ligands; n = 3 cultures each with 20 to 40 ROIs. (D) Summary of the fluorescence change measured in neurons expressing SST1.0 (D1) or CRF1.0 (D2) in response to a 2-hour continuous application of 1 μM SST-14 or 300 nM CRF, respectively; n = 4 cultures each with 20 to 40 ROIs. (E) Summary of normalized ΔF/F0 in HEK293T cells expressing SST1.0 (left) or CRF1.0 (right) in response to the indicated compounds applied: SST-14, CRF, UCN2, UCN3, CCK-8s, NPY, NTS, VIP, pituitary adenylate cyclase-activating polypeptide (PACAP), adrenocorticotropic hormone (ACTH), β-melanocyte-stimulating hormone (β-MSH), γ-MSH, growth hormone-releasing hormone (GHRH), oxytocin (OT), vasopressin (AVP), vosotocin, isotocin, glucagon, glucagon-like peptide (GLP-1), DAMGO, dynorphin A (DynA), enkephalin (Enk), goserelin, ghrelin, galanin, orexin A (OX-A), SP, neurokinin B (NKB), and bombesin (BB) were applied at 1 μM, while Glu, GABA, DA, norepinephrine (NE), serotonin (5-HT), histamine (HA), ATP, ADP, adenosine (Ado), teriparatide (TPTD), and taltirelin were applied at 10 μM (n = 4 wells containing 100 to 300 cells per well). (F and G) G protein and β-arrestin coupling were measured using the split-luciferase complementation assay (F1), a cyclic adenosine monophosphate (cAMP) reporter (F2), and the Tango assay (G1 and G2) in cells expressing either the wild-type peptide receptor (red), sensor (green), or no receptor (Ctrl; gray) in the presence of the indicated concentrations of the ligand; n = 3 wells each. AU, arbitrary units. (H) RNA-seq analysis shows that GRAB peptide sensors did not alter the cellular transcriptome. Comparisons of transcriptomes between cortex tissue expressing SST1.0 or EGFP-CAAX (H1) and between CRF1.0 or EGFP-CAAX (H2). Pearson's correlation coefficient analysis was used to evaluate the differential RNA expression.

Article Snippet: Where indicated, the following compounds were applied to the cells in Tyrode’s solution by bath application or a custom-made perfusion system: SST-28 (Anaspec), SST-14 (Anaspec), CCK-8s (Abcam), CCK-4 (Abcam), CRF (Anaspec), UCNI (MedChemExpress), UCNII (MedChemExpress), UCNIII (Abcam), NTS (Anaspec), NPY (Abcam), VIP (Anaspec), PACAP(1–38) (MedChemExpress), PACAP(1–27) (MedChemExpress), Orexin-B (GL Biochem), Substance P (Tocris), Ghrelin (Tocris), teriparatide [human parathyroid hormone-(1–34)] (MedChemExpress), Glu (Sigma-Aldrich), GABA (Tocris), DA (Sigma-Aldrich), NE (Tocris), 5-HT (Tocris), HA (Tocris), ATP (Tocris), ADP (MedChemExpress), Ado (Tocris), ACTH (MedChemExpress), β-MSH (MedChemExpress), γ-MSH (MedChemExpress), GHRH (Anaspec), OT (Anaspec), AVP (Tocris), vasotocin (MedChem-Express), isotocin (KS-V peptide), glucagon (GLPBIO), GLP-1 (MedChemExpress), DAMGO (Tocris), dynorphin A (Tocris), [Leu5]-enkephalin (MedChemExpress), goserelin (MedChemExpress), galanin (Tocris), orexin-A (Tocris), NKB (Tocris), bombesin (MedChemExpress), taltirelin (MedChemExpress), BIM23056 (Abcam), YM 022 (Tocris), NBI 27914 (Santa Cruz), antalarmin (Cayman), α-helical CRF (Tocris), SR142948 (Tocris), BIBO 3304 (Tocris), and PACAP(6–38) (Tocris).

Techniques: Cell Culture, Expressing, Membrane, Fluorescence, Luciferase, RNA Sequencing, RNA Expression

Journal: bioRxiv

Article Title: Tunable Rigid Spikes on Virus-Like Porous Silica Enable Mechanistically Controlled Nanovaccine Platforms

doi: 10.64898/2026.04.26.720861

Figure Lengend Snippet: (a) The flow chart of NPs-OVA immunized animals. (b) The serum levels of Anti-OVA IgG as measured by ELISA. (c) The serum levels of IL-2 measured by ELISA. (d, e) The ELISpot assay results for the secretion of IL-4 and IFN-γ by splenic cells. The P-value of less than 0.05 was considered statistically significant (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: Mouse IL-4 ELISA Kit (NBS EMC002.96), Mouse IFN-γ ELISA Kit (NBS EMC101g.96), Mouse IFN-γ ELISpotSASIC (HRP) (Mabtech 3321-2H), Mouse IL-4 ELISpotSASIC (HRP) (Mabtech 3311-2H), Tetramethylbenzidine (TMB) substrate for ELISpot (Mabtech 3651-10), OVA IgG ELISA Kit (Chondrex 3011), OVA IgM ELISA Kit (Chondrex 3017), and Low Endotoxin Ovalbumin from Chick Egg White (Chondrex 3022).

Techniques: Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal: bioRxiv

Article Title: Tunable Rigid Spikes on Virus-Like Porous Silica Enable Mechanistically Controlled Nanovaccine Platforms

doi: 10.64898/2026.04.26.720861

Figure Lengend Snippet: Results of subcutaneous injection of NPs-rEsxB in mice and subjected to lethal challenge with S. aureus. (a) The experimental flow chart for the lethal challenge. (b) The total serum rEsxB-specific IgG titer assay in NPs-rEsxB-immunized mice. (c, d) ELISA quantification of IL-4 and IFN-γ in mice. (e, f) The ELISpot assay results for the secretion of IL-4 and IFN-γ by splenic cells. (g, h) The survival rate of 1×LD100 S. aureus challenge experiment. (i, j, k) The survival rate of 2×LD100 S. aureus challenge experiment. All data are expressed as mean ± standard deviation (S.D.). The P-value of less than 0.05 was considered statistically significant (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: Mouse IL-4 ELISA Kit (NBS EMC002.96), Mouse IFN-γ ELISA Kit (NBS EMC101g.96), Mouse IFN-γ ELISpotSASIC (HRP) (Mabtech 3321-2H), Mouse IL-4 ELISpotSASIC (HRP) (Mabtech 3311-2H), Tetramethylbenzidine (TMB) substrate for ELISpot (Mabtech 3651-10), OVA IgG ELISA Kit (Chondrex 3011), OVA IgM ELISA Kit (Chondrex 3017), and Low Endotoxin Ovalbumin from Chick Egg White (Chondrex 3022).

Techniques: Injection, Titer Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Standard Deviation

Journal: PLoS ONE

Article Title: The Temporal and Hierarchical Control of Transcription Factors-Induced Liver to Pancreas Transdifferentiation

doi: 10.1371/journal.pone.0087812

Figure Lengend Snippet: Cultured adult human liver cells were infected with Ad-CMV-Pdx1 (1000 MOI), Ad-CMV-Pax4 (100 MOI) and Ad-CMV-Mafa (10 MOI) alone or in concert or with control virus ( Ad-CMV-β-gal , 1000 MOI), and pancreatic differentiation markers were examined six days later. (A) Immunofluorescence staining of human liver cells ectopically treated with a combination of three pTFs; Pdx1 (blue), Pax4 (green), and Mafa (red). Arrows indicate cells positive for all three pTFs. (B) Cultures were co-infected with the combined pTFs and with Ad-RIP-LUC (200 MOI), and Luciferase activity was measured. The results are expressed as Relative Light Unit (RLU)/mg protein. Each data point represents the mean ± SE of at least 2 independent experiments preformed in cells isolated from different donors, *p<0.05, **p<0.01 in comparison to control virus treated cells (n>4). (C) Immunofluorescence staining of treated human liver cells for insulin (red). Nuclei were stained with DAPI (blue), original magnification X20. The percent of insulin-positive cells was calculated by counting at least 500 positive cells from at least 2 independent experiments preformed in cells isolated from different donors. (D) Insulin (and or pro-insulin) secretion was measured by static incubation of the cells for 15 min at 2 and 17.5 mM glucose in KRB. n>12 in 5 independent experiments preformed in cells isolated from different donors, *p<0.05 comparing between triple infection and all other treatments.

Article Snippet: The antibodies used in this study were: anti-rabbit Pdx1, anti- goat Pdx1 (both 1∶1000 a generous gift from C.V. E. Wright), anti-human insulin, anti –human somatostatin (both 1∶100, Dako, Glostrup, Denmark), anti-Pax4 (1∶100; R&D Systems, Minneapolis, MN), anti-Mafa (1∶160; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Cell Culture, Infection, Virus, Immunofluorescence, Staining, Luciferase, Activity Assay, Isolation, Comparison, Incubation

Journal: PLoS ONE

Article Title: The Temporal and Hierarchical Control of Transcription Factors-Induced Liver to Pancreas Transdifferentiation

doi: 10.1371/journal.pone.0087812

Figure Lengend Snippet: Adult human liver cells were treated by the direct “hierarchical” sequential infection order (C, ). One single transcription factor (pTF) was omitted at a time and replaced by an identical MOI of Ad-CMV-β-gal . Pdx1 omission is indicated as (C-Pdx1), Pax4 omission is indicated as (C-Pax4), and Mafa omission is indicated as (C-Mafa). (A) Insulin promoter activation analysis, results are presented mean ± SE, * p<0.1, **p<0.05 compared to the direct “hierarchical” sequential infection order (C). n≥6 in 3 independent experiments preformed in cells isolated from different donors. (B) C-peptide secretion was measured by static incubation for 15 min at 2 and 17.5 mM glucose in KRB. *p<0.05, **p<0.01 compared to the direct “hierarchical” sequential infection order (C). n≥6 in 3 independent experiments. ( C–D ) Quantitative Real-Time PCR analysis for the transcription levels of pancreatic enzymes (C) and pTFs (D) . CT values are normalized to β-actin gene expression within the same cDNA sample. Results are presented as relative levels of the mean±SE compared to “hierarchy sequential infection” treated liver cells. *p<0.05, **p<0.01, n ≥6 in 3 independent experiments preformed in cells isolated from different donors.

Article Snippet: The antibodies used in this study were: anti-rabbit Pdx1, anti- goat Pdx1 (both 1∶1000 a generous gift from C.V. E. Wright), anti-human insulin, anti –human somatostatin (both 1∶100, Dako, Glostrup, Denmark), anti-Pax4 (1∶100; R&D Systems, Minneapolis, MN), anti-Mafa (1∶160; Santa Cruz Biotechnology, Inc., Santa Cruz, CA).

Techniques: Infection, Activation Assay, Isolation, Incubation, Real-time Polymerase Chain Reaction, Expressing