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: Cells

Article Title: Excessive Reactive Iron Impairs Hematopoiesis by Affecting Both Immature Hematopoietic Cells and Stromal Cells

doi: 10.3390/cells8030226

Figure Lengend Snippet: FeAS affects the expression of adhesion molecules on MS5 stroma cells and alters the profile of cytokine production. ( A ) After culture with or without FeAS, MS5 were stained as indicated and analyzed by fluorescence microscopy. Total ROS were detected with Red CC-1, intracellular labile iron with Calcein-AM, and nuclei with Hoechst 33,342. ( B ) After MS5 cells were cultured with or without FeAS for 48 h, cells were stained with the indicated Abs. Representative flow cytometry histograms from three independent experiments are shown. ( C ) Conditioned media were analyzed with a cytokine array and the accompanying kit reagents and protocol. Conditioned media from FeAS-treated (10 or 50 μM) or untreated MS5 cells (CTL) was collected after 48 h. of culture. Medium supplemented with 10% FBS αMEM was used as a negative control (NC). The assay is arranged in a 14 × 10 grid (Array 3) and a 12 × 8 grid (Array 4), with the configuration shown in . Summarized densitometry data (mean ± SD) from three independent experiments is shown in the right panel (* p < 0.05). ( D ) MS5 cells were cultured in the presence or absence of FeAS for 48 h, and co-cultured with LSKs with or without FeAS for another 48 h and the number of cobblestone-like areas seen in each well were counted under light microscopy. Summarized data (mean ± SD) from two independent experiments is shown (* p < 0.05). Unt-FeAS: FeAS was present in the culture medium only when MS5 cells were co-cultured with LSK; FeAS-CTL: FeAS was present in the culture medium when MS5 cells were cultured alone but not when they were co-cultured with LSK; FeAS-FeAS: FeAS was present in the culture medium in both, when MS5 cells were cultured alone and when they were co-cultured with LSK cells; and Unt-CTL: FeAS was not present in the culture medium neither when MS5 cells were cultured alone nor when they were co-cultured with LSK.

Article Snippet: Cells stained with Calcein-AM, RedCC-1, and Hoechst 33,342 were analyzed with a Zeiss Laser Scanning microscopy system (Zeiss LSM5 PASCAL, Obercochen, Germany).

Techniques: Expressing, Staining, Fluorescence, Microscopy, Cell Culture, Flow Cytometry, Negative Control, Light Microscopy

Journal: eLife

Article Title: Zebrafish fin regeneration involves generic and regeneration-specific osteoblast injury responses

doi: 10.7554/eLife.77614

Figure Lengend Snippet: ( A ) Experimental scheme indicating EdU intraperitoneal (IP) injections used to label cycling osteoblasts harvested at different time points for the data presented in panels B–D. ( B ) Proliferation timeline of bglap :GFP+ osteoblasts in segment –1 as determined by EdU incorporation. N (experiments)=1, n (fins)=4 (0 hpa), 10 (1 day post amputation [dpa]), 9 (2 dpa); n (rays)=14 (0, 2 dpa), 10 (1 dpa). Error bars represent 95% CI. Kruskal-Wallis test. ( C ) Proliferation of bglap :GFP+ osteoblasts in segment –2, segment –1, and segment 0 at 2 dpa. N (experiments)=1, n (fins)=9, n (rays)=14. Error bars represent 95% CI. Dunn’s test. ( D ) bglap RNAscope expression levels of EdU+ cells in segment –1 at 2 dpa. bglap -expressing osteoblasts were categorised based on RNAscope signal intensity into ‘high’ (yellow), ‘medium’ (cyan), and ‘low’ (red) using a look-up table, with ‘high’ threshold corresponding to expression levels in segment –2. N (experiments)=1, n (fins)=9, n (rays)=12, n (cells)=62. Error bars repesent SD. Scale bar, 10 µm.

Article Snippet: To analyse cell proliferation, 5-ethynyl-2′-deoxyuridine (EdU)-Click 647 kit (baseclick GmbH BCK-EdU647) was used.

Techniques: RNAscope, Expressing