Journal: Cell Reports Medicine

Article Title: Modulation of lipid nanoparticle-formulated plasmid DNA drives innate immune activation promoting adaptive immunity

doi: 10.1016/j.xcrm.2025.102035

Figure Lengend Snippet:

Article Snippet: For SARS-CoV-2 spike binding ELISAs in B–7E, assays were performed as above, except for coating with the following proteins where indicated: SARS-CoV-2 wild type spike RBD (Cat# 40592-V08H, Sino Biologicals), SARS-CoV-2 D614G spike (Cat# 40589-V08H8, Sino Biologicals), SARS-CoV-2 B.1.617.2 spike (Cat# 40589-V08H10, Sino Biologicals), SARS-CoV-2 B.1.1.529 spike (Cat# 40589-V08H26), or SARS-CoV-2 BA.2 spike (Cat# 40589-V08H28, Sino Biologicals).

Techniques: Virus, Recombinant, Lysis, Reporter Gene Assay, Cell Stimulation, Electron Microscopy, Luminex, Enzyme-linked Immunospot, Luciferase, Plasmid Preparation, Software, Synthesized

Journal: American Journal of Cancer Research

Article Title: Inhibition of S-adenosylhomocysteine hydrolase decreases cell mobility and cell proliferation through cell cycle arrest

doi:

Figure Lengend Snippet: Information about antibodies used to western blotting analysis

Article Snippet: Used-antibodies were organized in . table ft1 table-wrap mode="anchored" t5 caption a7 Name Cat No. Company AHCY ab134966 Abcam Ras (27H5) #3339 Cell signaling Phospho-B-Raf (Ser445) #2696 Cell signaling B-Raf (55C6) #9433 Cell signaling Phospho-C-Raf (Ser259) #9421 Cell signaling C-Raf #9422 Cell signaling Phospho-MEK1/2 (Ser221) (166F8) #2338 Cell signaling MEK1/2 (L38C12) #4694 Cell signaling phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) #4377 Cell signaling p44/42 MAPK (Erk1/2) #9102 Cell signaling cyclinD1 (92G2) #2978 Cell signaling CDK6 (DCS83) #3136 Cell signaling cyclinE (HE12) #4129 Cell signaling CDK2 sc-163 Santa cruz cyclinB1 (D5C10) XP ® #12231 Cell signaling phospho-cdc2 (Tyr15) (10A11) #4539 Cell signaling cdc2 (POH1) #9116 Cell signaling p21 waf1/cip1 (DCS60) #2946 Cell signaling chk2 NB100-500 Novus Bio ATM NB100-309 Novus Bio phospho-MDM2 (Ser166) sc-293105 Santa cruz MDM2 (SMP14) sc-965 Santa cruz phospho-p53 (Ser15) sc-101762 Santa cruz p53 (DO-1) sc-126 Santa cruz Gadd45α (D17E8) #4632 Cell signaling 14-3-3ζ/δ (D7H5) #7413 Cell signaling cytoskeletal actin A300-291A Bethyl goat anti-rabbit IgG, pAb ADI-SAB-300-J Enzo goat anti-mouse IgG, pAb ADI-SAB-100 Enzo anti-biotin HRP-linked Ab #7075 Cell signaling Open in a separate window Information about antibodies used to western blotting analysis Cell proliferation assay Approximately 1 × 10 4 cells were seeded per well in 12-well cell culture plates in 10% FBS-DMEM.

Techniques: Western Blot

Journal: bioRxiv

Article Title: The β-cell primary cilium is an autonomous Ca 2+ compartment for paracrine GABA signalling

doi: 10.1101/2021.08.16.456473

Figure Lengend Snippet: A. Immunofluorescence staining of a mouse islet showing acetylated tubulin in magenta, GABA-B1 receptors in green and the nuclei in blue (Dapi). B. Immunofluorescence staining of a mouse islet showing acetylated tubulin in green and GABA-B1 receptors in blue (anti-goat) and magenta (anti-rabbit). C. Immunofluorescence staining of a mouse islet showing acetylated tubulin in magenta and GABA-A receptors in green. D. Representative images showing the cilia localization of GABA-B1 receptors (magenta) to the primary cilium (green) in MIN6 cells, MIN6 pseudo-islets, mouse islet cells, mouse islet cells transduced with Ad-Smo-GCaMP5G-mCh and human islet cells. At the bottom is shown a cilium from a mouse islet stained for acetylated tubulin (green) and GABA-B2 receptors (magenta). E. Quantifications of the cilia distribution of GABA-B1 receptors in the different cell preparations. F. STED images of a mouse islet cell cilia stained for acetylated tubulin (green) and GABA-B1 receptors (magenta). G. Center-of-mass of acetylated tubulin and GABA-B1 receptors from “straightened” cilia (example above) of mouse islets (n=39, cilia from 25 islets). H. Distribution of GABA-B1 receptors at the base of the cilium quantified from STED images. Notice that the receptors localize peripheral to the microtubule core of the cilium (means ± S.E.M. for 39 cilia from 25 islets). I. Center-of-mass analysis of mouse islet cilia stained for acetylated tubulin and GABA-B1 receptors under control conditions and following a 10 min incubation with 10 nM GABA. Notice that the addition of GABA results in a shift in the center of mass towards more distal regions of the cilium (n=39, control and n=27, GABA cilia from at least 25 islets each, **** P<0.0001, One Way ANOVA with Multiple Comparisons).

Article Snippet: The following antibodies were used in the study: acetylated tubulin (T7451 Sigma, host: mouse, 1:500), IFT88 (F41236 NSJ Bioreagents, host: rabbit, 1:200), SSTR3 (E-AB-1607 Elabscience, host: rabbit, 1:200), Patched (ab53715 Abcam, host: rabbit, 1:200), Smoothened (ab113438 Abcam, host: rabbit, 1:200), GABA-B1 receptor (NBP1-52389 Novus Biologicals, host: goat, 1:200.

Techniques: Immunofluorescence, Staining, Transduction, Control, Incubation

Journal: bioRxiv

Article Title: The β-cell primary cilium is an autonomous Ca 2+ compartment for paracrine GABA signalling

doi: 10.1101/2021.08.16.456473

Figure Lengend Snippet: A. TIRF microscopy recordings of GCaMP5G fluorescence in the soma and cilium of MIN6 cells exposed to 1 nM GABA. Notice that the addition of GABA induces a rise of cilia Ca 2+ without affecting the Ca 2+ concentration in the soma. B. TIRF microscopy recordings of GCaMP5G fluorescence in the soma and cilium of MIN6 cells exposed to 10 nM GABA. Notice that the addition of GABA induces a rise of cilia Ca 2+ without affecting the Ca 2+ concentration in the soma. C. GABA (1 and 10 nM) and the GABA-B receptor agonist Baclofen induces increases in the cilia Ca 2+ concentration in MIN6 cells (n=10 cells for 1 nM GABA; n=13 cells for 10 nM GABA; n=7 cells for Baclofen; * P<0.05; ** P<0.01; *** P<0.001; Student’s paired t-test). D. TIRF microscopy recordings of cilia Ca 2+ concentration changes in intact mouse islets expressing Smo-GCaMP5G-Ch. Picture on top shows the distribution of cilia within the islet, with cilia exhibiting at least one spontaneous Ca 2+ flash during the recording time highlighted in color. In the middle are shown Ca 2+ recordings from the individual cilia within the islet during addition of the GABA-B receptor agonist Baclofen. The histogram at the bottom shows the overall cilia Ca 2+ activity of the whole islet. E. Histogram showing the number of cilia Ca 2+ flashes following the addition of baclofen (n=7 islets). F. Histogram showing the number of cilia Ca 2+ flashes following the addition of vigabatrin (n=13 islets). G. Histogram showing the number of cilia Ca 2+ flashes following the addition and washout of picorotoxin and CGP-35348 (n=8 islets). H. Histogram showing the number of cilia Ca 2+ flashes following preincubation with DMSO (control) and vigabatrin for 5 h (n=5 islets for both conditions). I. Quantifications of the effect of baclofen, vigabatrin, picorotoxin+CGP-35348 and long-term vigabatrin treatment on cilia Ca 2+ flashes. Statistical significance was assessed using the Friedman test followed by multiple comparisons.

Article Snippet: The following antibodies were used in the study: acetylated tubulin (T7451 Sigma, host: mouse, 1:500), IFT88 (F41236 NSJ Bioreagents, host: rabbit, 1:200), SSTR3 (E-AB-1607 Elabscience, host: rabbit, 1:200), Patched (ab53715 Abcam, host: rabbit, 1:200), Smoothened (ab113438 Abcam, host: rabbit, 1:200), GABA-B1 receptor (NBP1-52389 Novus Biologicals, host: goat, 1:200.

Techniques: Microscopy, Fluorescence, Concentration Assay, Expressing, Activity Assay, Control