Journal: Scientific Reports

Article Title: Mitochondrial behaviour throughout the lytic cycle of Toxoplasma gondii

doi: 10.1038/srep42746

Figure Lengend Snippet: (a) Fluorescence micrograph of intracellular (In) and extracellular (out) populations of T. gondii tachyzoites taken utilizing the signal from 215430-YFP (green in the merge with the brightfield, top panels, and in the bottom panels). Bars 5 μm. (b) Three representative images of each of the observed shapes of mitochondria in extracellular parasites and their classification. Each example shows the fluorescent signal image on the left and the merge of fluorescence and brightfield on the right. The color-coding of the frame (lasso – green; sperm-like – orange; collapsed – red) is maintained throughout all the figures. (c) Proportions of the morphologies scored in intracellular parasites (Intracellular, 776 parasites, over 6 independent experiments); in parasites mechanically released from host cell into full growth medium (Full, 864 parasites, over 9 independent experiments) or into diluted growth medium (12%, 653 parasites, over 2 independent experiments) immediately after release; and in parasites induced to egress by 2 μM ionomycin (Ionomycin, 1177 parasites, over 6 independent experiments) immediately after release. Error bars are standard deviation. (d) Super-resolution microscopy images of the mitochondrial lasso morphology in intracellular (left) and the three main mitochondrial morphologies observed in extracellular: lasso, sperm-like and collapsed (right) shown as projection of all Z stacks (top) and as 3D reconstruction (bottom). 215430 - green. DAPI - blue. Bar 2 μm.

Article Snippet: Images were taken every 10 seconds for a total of 5 minutes and 2 μM ionomycin (Santa Cruz Biotechnology) was added after 2–4 time points were imaged.

Techniques: Fluorescence, Standard Deviation, Super-Resolution Microscopy

Journal: eLife

Article Title: Mechanically stimulated ATP release from murine bone cells is regulated by a balance of injury and repair

doi: 10.7554/eLife.37812

Figure Lengend Snippet: ( A ) Confocal images of CB-OB loaded with quinacrine and MANT-ATP, . ( B ) Changes in quinacrine-loaded CB-OB treated with ionomycin ( left ) or stimulated with micropipette ( right , region of interest in white). ( C ) Vesicular release event in CB-OB ( top ) coincides with a sudden drop in granular quinacrine fluorescence ( bottom ), .( D ) Kinetics of vesicular release from CB-OB stimulated by micropipette at 0 s, n = 37 stimulated cells. ( E–H ) Quinacrine- ( E, F ) or Fura2- ( G, H ) loaded CB-OB were pre-treated (10 min) with vehicle, ionomycin (iono) or NEM or placed in [Ca 2+ ]-depleted physiological solution, and micropipette-stimulated when indicated (+). Vesicular density ( E ) and cumulative release ( F ), secondary responsiveness ( G ) and [Ca 2+ ] response activation rates ( H ) were determined, n = 7–37 primary cells. ( I ) tFSS was applied by replacing 50% media volume n times. ( J, K ) ATP released per cell ( left axis ) or as percent of cellular ATP content ( right axis ) was measured following CB-OB stimulation by tFSS ( J , black dashed line : rational function fit; red dashed line: corresponding asymptote) or after indicated pre-treatments followed by tFSS ( K , 10x media displacements, +), n = 6–8 independent cultures. For , means ± SEM, *significance compared to vehicle ( E–H ), basal ATP release ( J ) or to tFSS-stimulated vehicle ( K ) by ANOVA. Source data for is provided in . 10.7554/eLife.37812.017 Figure 3—source data 1.

Article Snippet: Chemical compound, drug , Ionomycin calcium salt; Iono , Tocris Bioscience , Cat. 1704 , calcium ionophore, 100 μM.

Techniques: Fluorescence, Activation Assay

Journal: eLife

Article Title: Mechanically stimulated ATP release from murine bone cells is regulated by a balance of injury and repair

doi: 10.7554/eLife.37812

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , Ionomycin calcium salt; Iono , Tocris Bioscience , Cat. 1704 , calcium ionophore, 100 μM.

Techniques: In Vivo, Software, Cell Culture, In Vitro, Viability Assay, Staining, Luciferase, ATP Bioluminescent Assay, ALP Assay, Modification

Journal: Med (New York, N.y.)

Article Title: A booster dose enhances immunogenicity of the COVID-19 vaccine candidate ChAdOx1 nCoV-19 in aged mice

doi: 10.1016/j.medj.2020.12.006

Figure Lengend Snippet: ChAdOx1 nCoV-19 induces a Th1-dominated CD4 cell response (A) tSNE/FlowSOM analyses of CD4 + T cells from 3-month-old (3mo) mice 7 days after immunization with ChAdOx1 nCoV-19 or PBS; on the heatmap, red indicates high expression, and yellow indicates low expression. (B and C) Heatmaps of the manually gated CD4 + Foxp3 – (B) and Foxp3 + CD4 + (C) T cell populations indicated at 7, 14, and 21 days after immunization in the iliac lymph node (right) and spleen (left), the gating strategy for these populations is shown in Figure S2 . Here, the frequency of each cell subset in each ChAdOx1 nCoV-19-immunized mouse has been expressed as the log2 fold change over the average frequency in PBS-immunized mice (n = 5). (D–G) Bar charts showing the number of CD69 + CD62L + CD44 – CD4 + Foxp3 – (D), Ki67 + CD4 + Foxp3 – (E) CXCR3 + non-Tfh cells (F) and CXCR3 + Th1-like Treg cells (G) CD4 + cells in the iliac lymph node of ChAdOx1 nCoV-19 or PBS-immunized mice, at the indicated time points post-immunization. (H) Analysis of cytokine production 6 h after PdBu/ionomycin stimulation of iliac lymph node cells from 3-month-old mice 7 days after immunization with ChAdOx1 nCoV-19 or PBS. (I and J) Stacked bar plots show the number of CD4 + Foxp3 – cells singly or co-producing IFN-γ, IL-2, or TNF-α in the iLN 7 days after immunization (I) or spleen at the days post-immunization (J); 6 h after restimulation with SARS-CoV-2 peptide pools, each bar segment represents the mean and the error bars the standard deviation. In (D)–(G), the bar height corresponds to the mean and each circle represents one biological replicate. p values are calculated using a Student’s t test with Holm-Sidak multiple testing correction.

Article Snippet: Ionomycin calcium salt , Tocris Bioscience , Cat# 1704/1.

Techniques: Expressing, Standard Deviation

Journal: Med (New York, N.y.)

Article Title: A booster dose enhances immunogenicity of the COVID-19 vaccine candidate ChAdOx1 nCoV-19 in aged mice

doi: 10.1016/j.medj.2020.12.006

Figure Lengend Snippet: ChAdOx1 nCoV-19 induces a CD8 T cell response (A) tSNE/FlowSOM analyses of CD8 + T cells from 3-month-old (3mo) mice 7 days after immunization with ChAdOx1 nCoV-19 or PBS; on the heatmap red indicates high expression, and yellow indicates low expression. (B) Heatmap of the manually gated CD8 T cell populations indicated at 7, 14, and 21 days after immunization in the iliac lymph node and spleen; the gating strategy for these populations is shown in Figure S2 . The frequency of each cell subset in each ChAdOx1 nCoV-19-immunized mouse has been expressed as the log2 fold change over the average frequency in PBS-immunized mice (n = 5). Crossed boxes indicate that there were none of that cell type for that mouse. (C–F) Bar charts showing the number of Ki67 + (C), antigen-experienced CD44 + CD62L – (D), CXCR3 + (E), and PD-1 + CD44 + (F) CD8 cells in the iliac lymph node of ChAdOx1 nCoV-19 or PBS-immunized mice, at the indicated time points post-immunization. (G) Analysis of cytokine production 6 h after PdBu/ionomycin stimulation of iliac lymph node cells from 3-month-old mice 7 days after immunization with ChAdOx1 nCoV-19 or PBS. (H and I) Stacked bar plots show the number of CD8 + cells singly or co-producing Granzyme B, IFN-γ, IL-2, or TNF-α 6 h after restimulation with SARS-CoV-2 peptide pools, in the iLN 7 days after immunization (H) or spleen 7, 14, and 21 days after immunization (I); each bar segment represents the mean and the error bars the standard deviation. In (C)–(F), the bar height corresponds to the mean, and each circle represents one biological replicate. p values are calculated using a Student’s t test with Holm-Sidak multiple testing correction.

Article Snippet: Ionomycin calcium salt , Tocris Bioscience , Cat# 1704/1.

Techniques: Expressing, Standard Deviation

Journal: Med (New York, N.y.)

Article Title: A booster dose enhances immunogenicity of the COVID-19 vaccine candidate ChAdOx1 nCoV-19 in aged mice

doi: 10.1016/j.medj.2020.12.006

Figure Lengend Snippet:

Article Snippet: Ionomycin calcium salt , Tocris Bioscience , Cat# 1704/1.

Techniques: Blocking Assay, Antibody Labeling, Activation Assay, Marker, Control, Purification, Conjugation Assay, Virus, Recombinant, Staining, Transfection, Vaccines, Luciferase, Gene Expression, Expressing, Plasmid Preparation, Software