Journal: bioRxiv

Article Title: Lymph node resident memory T cells retain effector capabilities by evading lung resident memory dysfunction

doi: 10.64898/2026.02.03.703569

Figure Lengend Snippet: (A) Naive CD90.1+ OT-I T cells were transferred into naive B6 mice before intranasal infection with influenza expressing SIINFEKL (PR8-OVA). Expression of CD69 and CD103 was measured 21 days post infection on extravascular cells. (B) Quantification of CD69 + CD103 + OT-I T cells. (C) Naïve CD90.1 + OT-I Nur77-GFP T cells were transferred into mice one day before intranasal PR8-OVA infection. Nur77-GFP expression was evaluated 3-14 days later by extravascular OT-I T cells. (D)Experimental setup for (E-G). Flow cytometry plot (E) of CD90.1 + OT-I T cells isolated from indicated locations. (F-G) Longitudinal labeling of cells and quantification. (H) Experimental setup for (I-J). (I) Labeling after intravenous and intraperitoneal injection of OT-I T cells or naïve endogenous T cells. (J) Expression of CD69 and CD103. Naïve= CD8 + CD90.1 - CD44 low . Each point represents a mouse and N=2-5 mice per experiment. Data are representative (A) or are pooled from at least two experiments (n=2–4/group), except for day 113 in (B), which reflect a single experiment.

Article Snippet: Measurements were obtained in Flow Cytometry (FACS) Staining Buffer (Rockland, MB-086-0500) at room temperature for about 30 minutes per sample and channels were flushed with buffer in between samples.

Techniques: Infection, Expressing, Flow Cytometry, Isolation, Labeling, Injection

Journal: bioRxiv

Article Title: Lymph node resident memory T cells retain effector capabilities by evading lung resident memory dysfunction

doi: 10.64898/2026.02.03.703569

Figure Lengend Snippet: (A)Chromatin accessibility with gene expression (right) and linked chromatin peaks (bottom) at the Gzmb locus. (B) Protein expression of GranzymeB as measured by flow cytometry. (C-D) Same as (A-B) but for the Pdcd1 locus. Protein expression on OT-I T cells is 49 days post infection where each dot represents one mouse. (J and L) Data are from one experiment and representative of at least two experiments with N=3. (E-F) Surface expression of PD-1 and TIM3 at 7 days post infection. (G) Transcription factor motif footprint analysis from dataset. (H) Inferred eRegulons identified by SCENIC+ across memory T cells after influenza infection. (I-K) Legend (left) for network plots illustrating inferred transcription factor relationships to target genes. Transcription factor and gene relationship is denoted by color of line represents activation (red) or repression (blue).

Article Snippet: Measurements were obtained in Flow Cytometry (FACS) Staining Buffer (Rockland, MB-086-0500) at room temperature for about 30 minutes per sample and channels were flushed with buffer in between samples.

Techniques: Gene Expression, Expressing, Flow Cytometry, Infection, Activation Assay

Journal: Scientific Reports

Article Title: A Bioluminescent Cell Assay to Quantify Prion Protein Dimerization

doi: 10.1038/s41598-018-32581-1

Figure Lengend Snippet: PrP-NGLuc and PrP-CGLuc expression in RK13-DC cells results in bioluminescence. ( a ) Western blot analysis with the Sha31 antibody against PrP shows that RK13-DC cells stably express PrP-NGLuc and PrP-CGLuc from a bicistronic expression vector (lanes 3 and 4). Non-transfected RK13 cells do not express PrP and do not show a detectable signal for PrP (lanes 1 and 2), whereas N2a cells show the characteristic signal for wild-type PrP (lanes 5 and 6). Deglycosylation of cell lysates with peptide-N-glycosidase F (PNGase F, lanes 2, 4, and 6) resulted in lower molecular weight bands suggesting that mature PrP-NGLuc and PrP-CGLuc are properly glycosylated in RK13-DC cells as PrP is in N2a cells. Detection of tubulin on the same blot served as a loading control. Additional lanes were excised for presentation purposes. ( b ) Western blot analysis with the Sha31 antibody against PrP shows that treatment of RK13-DC cells with increasing amounts of phosphoinositide phospholipase C (PI-PLC) increases the amount of detectable protein released into cell culture medium suggesting that mature PrP-NGLuc and PrP-CGLuc are anchored to the outer cell membrane by GPI. Additional lanes were excised for presentation purposes. ( c ) Densitometric quantification of the PrP signal from three different experiments as shown in ( b ). ( d ) Life cell immunofluorescence staining with the Sha31 antibody to PrP (left panels) shows that PrP-NGLuc and PrP-CGLuc are expressed in RK13-DC cells (top row), similar to wild-type PrP in N2a cells (bottom row). Bright-field microscopy (centre panels) shows the contours of the imaged cells. An overlay of fluorescence and bright-field images (right panels) shows that PrP-NGLuc, PrP-CGLuc, and PrP are located at the cell membrane. Bar = 20 µm. ( e ) The bioluminescence measured from RK13-DC cells expressing both PrP-NGLuc and PrP-CGLuc was more than 10-fold above background levels measured from RK13 cells stably expressing only PrP-NGLuc or PrP-CGLuc, or both NGLuc and CGLuc together each lacking the PrP moiety. A functional luciferase consisting of its N- and C-terminal halves was only formed in RK13-DC cells where dimers between the PrP moieties of PrP-NGLuc and PrP-CGLuc could form. In comparison to PrP-NGLuc and PrP-CGLuc in RK13-DC cells, bioluminescence of full-length GLuc in RK13 cells was almost 17-fold higher. In ( c , e ) error bars indicate SD.

Article Snippet: Additional lanes were excised for presentation purposes. ( c ) Densitometric quantification of the PrP signal from three different experiments as shown in ( b ). ( d ) Life cell immunofluorescence staining with the Sha31 antibody to PrP (left panels) shows that PrP-NGLuc and PrP-CGLuc are expressed in RK13-DC cells (top row), similar to wild-type PrP in N2a cells (bottom row).

Techniques: Expressing, Western Blot, Stable Transfection, Plasmid Preparation, Transfection, Molecular Weight, Control, Cell Culture, Membrane, Immunofluorescence, Staining, Microscopy, Fluorescence, Functional Assay, Luciferase, Comparison