Journal: Cell

Article Title: RIPK3 restricts viral pathogenesis via cell death-independent neuroinflammation

doi: 10.1016/j.cell.2017.03.011

Figure Lengend Snippet: Key Resource Table

Article Snippet: Cells were stained using a primary anti-mCherry antibody (Rockland) for 1 hour and goat anti-rabbit AlexaFluor 594 secondary antibody (ThermoFisher) for 15 minutes, both at room temperature.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software

Journal: Heliyon

Article Title: Median raphe glutamatergic neuron-mediated enhancement of GABAergic transmission and suppression of long-term potentiation in the hippocampus

doi: 10.1016/j.heliyon.2024.e38192

Figure Lengend Snippet: MRN VGLUT3 neuron-mediated glutamatergic transmission in SR/SLM 5-HT3aR neurons. A) A schematic of AAV injection into the MRN. The left panel shows an image of the MRN from VGLUT3 Cre/5-HT3aR-EGFP mouse which received an AAV-EF1a-double floxed-hChR2(H134R)-mCherry-WPRE-HGHpA injection into the MRN. Scale 150 μm. B) An image of the hippocampus from MRN VGLUT3 ChR2/5-HT3aR-EGFP mouse. The inset shows EGFP-expressing SR/SLM 5-HT3aR neurons and mCherry-expressing MRN VGLUT3 axons. Scale 300 μm/25 μm (inset). C) The upper panel shows a schematic for recording MRN VGLUT3 neuron-mediated transmission in SR/SLM 5-HT3aR neurons. The bottom panel shows example traces for MRN VGLUT3 neuron-mediated EPSCs in SR/SLM 5-HT3aR neurons without drug application, in the presence of tetrodotoxin (TTX, 1 μM), TTX+4-aminopyridine (4-AP, 100 μM) and TTX+4-AP + DNQX (10 μM). Scale 50 pA/10 ms. The blue line indicates light application. D) EPSC amplitude in SR/SLM 5-HT3aR neurons before the drug application, in the presence of TTX, TTX+4-AP, and TTX+4-AP + DNQX (6 neurons from 3 female mice and 6 neurons from 3 male mice). F (1,11) = 22.01, P = 0.001. Data are presented as mean ± SEM. E) EPSC amplitude in female (17 neurons/5 mice) and male (16 neurons/5 mice) groups. Female and male groups did not show statistically significant difference in EPSC amplitude (P = 0.8). The horizontal line in each group represents the mean and the vertical line represents SEM. F) The delay between the onset of the light stimulation and the onset of EPSCs in female (17 neurons/5 mice) and male (16 neurons/5 mice) groups. Female and male groups did not show a statistically significant difference in the delay between the onset of light stimulation and the onset of EPSCs (P = 0.77). n.s. not statistically significant.

Article Snippet: The slices were then incubated in mCherry polyclonal antibody (Rockland Immunochemicals, Cat. No. 600401P16) or Anti-GFP antibody (Abcam, Cat. No. ab13970, RRID: AB_300798 ) at a dilution of 1:1000 in 0.3 % Triton-X/TBS for 24 h at 4 °C [ , ].

Techniques: Transmission Assay, Injection, Expressing

Journal: The EMBO Journal

Article Title: Hybrid endosomal coats contain different classes of sorting nexins

doi: 10.1038/s44318-026-00716-0

Figure Lengend Snippet: ( A ) Colocalization of Snx3 and SNX-BARs. Wild-type or vps35Δ cells co-expressing Vps5 yomCherry and Vps17 mNG , or Vps5 yomCherry and mNG Snx3 were analysed by a z-series of confocal images taken at a z-distance of 0.3 µm. Maximum projections were generated in ImageJ. Scale bar: 5 μm. ( B ) Pearson correlation coefficient (PCC) values were calculated using the JACoP plugin in the ImageJ software for a total of >80 cells from three independent experiments. See Fig. for biological replicate variability. Bars represent the mean values and standard deviation. A two-tailed unpaired t test was used to evaluate significance. **** P < 0.0001; *** P < 0.001; ** P < 0.01 and * P < 0.1. ( C ) Immunoprecipitation of mCherry-tagged Vps35 wt or Vps35 QGRE . Logarithmically growing vps35Δ cells expressing Vps35 mCherry (WT) or Vps35 QGRE-mCherry (Mut) from an integrative plasmid, and genomically tagged Snx3 V5 and Vps5 HA , were lysed in detergent. Proteins were pulled down with an affinity matrix to mCherry and analysed by SDS-PAGE and Western blotting against the indicated tags. ( D ) Quantification of the amount of Vps5 and Snx3 pulled down by mCherry-labelled Vps35 wt or Vps35 QGRE . The ratios between the signals for Vps35 and Snx3 or Vps5 were calculated. For Vps35 WT the ratio was set to 1 as a reference. Mean and standard deviation from five independent experiments are shown. A two-tailed unpaired t test was used to evaluate significance. **** P < 0.0001; *** P < 0.001; ** P < 0.01 and * P < 0.1. ( E ) Snx3 V5 was pulled down in an experiment as in ( C ), using vps35Δ cells expressing Vps35 mCherry (WT) or Vps35 QGRE-mCherry (Mut) from a plasmid and genomically tagged Snx3 V5 and Vps5 HA in the indicated combinations. ( F ) Quantification of Vps35 mCherry and Vps5 HA pulled down by Snx3 V5 . The ratios between the signals for Snx3 V5 and Vps35 mCherry or Vps5 HA were calculated. For Vps35 wt , the ratio was set to 1 as a reference. Mean and standard deviation from three to five independent experiments are shown. A two-tailed unpaired t test was used to evaluate significance. **** P < 0.0001; *** P < 0.001; ** P < 0.01 and * P < 0.1. .

Article Snippet: mCherry Polyclonal antibody , Chromotek , 26765-1-AP.

Techniques: Expressing, Generated, Software, Standard Deviation, Two Tailed Test, Immunoprecipitation, Plasmid Preparation, SDS Page, Western Blot

Journal: The EMBO Journal

Article Title: Hybrid endosomal coats contain different classes of sorting nexins

doi: 10.1038/s44318-026-00716-0

Figure Lengend Snippet: Logarithmically growing vps35Δ cells expressing genomically tagged Vps10 mNG were transformed with integrative plasmids expressing Vps35 mCherry (WT), Vps35 QGRE-mCherry , or nothing. The cells were logarithmically grown overnight and analysed by spinning disc microscopy. Scale bar: 5 µm.

Article Snippet: mCherry Polyclonal antibody , Chromotek , 26765-1-AP.

Techniques: Expressing, Transformation Assay, Microscopy