Journal: Journal of Lipid Research

Article Title: Hepatoprotective drug screening identifies daclatasvir, a promising therapeutic candidate for MASLD by targeting PLIN2

doi: 10.1016/j.jlr.2025.100835

Figure Lengend Snippet: Daclatasvir treatment ameliorates liver damage and fibrosis in MCD diet-induced MASH mouse model. A: Schematic illustration of the experimental workflow for the daclatasvir treatment strategy in a methionine-choline deficient (MCD) diet-induced MASH mouse model. B: Liver weight, LW/BW, and LW/BMI were recorded for MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. C: Representative images of H&E, PSR, and CD11b staining of liver samples from vehicle or daclatasvir treated mice at 4 weeks of MCD diet administration. Scale bar, 200 μm for H&E and PSR; 100 μm for CD11b. D: Quantitative results for PSR and CD11b staining shown in (C). E: Enzyme activities of serum hepatic transaminases in MCD diet-fed mice after 3 weeks of treatment with either vehicle or daclatasvir. F–H: The mRNA expression of genes associated with lipid catabolism (F), inflammation (G), and fibrosis (H) in liver tissues from MCD diet-induced MASH mice following 3 weeks of daclatasvir or vehicle treatment. For (B–H), n = 6 mice per group. Data are presented as mean ± SD; ∗ P < 0.05, ∗∗ P < 0.01; Student’s t test for statistics.

Article Snippet: The sections were incubated with a rabbit anti-CD11b primary antibody (BM3925, Boster) overnight at 4°C, followed by incubation with a goat anti-rabbit fluorophore-conjugated secondary antibody (A-11036, Invitrogen).

Techniques: Staining, Expressing

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: (Α) Histology of polyps from APC Δ468 mouse. (a–b) CEA stained paraffin sections counterstained with Gill's II Hematoxylin; (c&d) methylene blue staining; arrows point to (a) granulocytes, (b) mast cells, (c) plasma cells, (d) lymphocytes. (B) Immuno-fluorescence of polyps from APC Δ468 mice. Cryosections were stained with (a) DAPI, (b) CD11b-AlexaFluor 488, and (c) Gr1-AlexaFluor 594. Arrows point to polyp, and arrowheads to the adjacent healthy villus; note accumulation of CD11b + cells and/or Gr1 + cells in the polyps. (C) FACS analysis of leukocytes prepared from micro-dissected polyps, and from adjacent tissue (n = 4), and of intestinal tissue from age-matched healthy control mice (n = 3). Mean values and SEM are shown for frequencies of CD11b + and of CD11b + Gr1 + cells in 6 month-old mice.

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: Staining, Clinical Proteomics, Fluorescence, Control

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: (A) FACS analyses of pro-inflammatory cells. (a) Exemplar FACS analyses of mononuclear cells prepared from the intestine of (a&c) a 5 month-old APC Δ468 mouse and (b&d) an age matched wt C57BL/6J mouse; note increases in the frequencies of (a) CD11b + Gr1 + and (c) CD11b + F4/80 + myeloid type cells in the polyposis intestine, compared to wt tissue (b&d respectively). (B) Summary of FACS analysis of MNCs from wt (black bars) or APC Δ468 (open bars) intestine, showing mean frequencies and absolute numbers of cells with SEM values; n = 6 for APC Δ468 , n = 3 for wt control mice, per age group.

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: Control

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: Cryosections of ProSense-680 in vivo stained intestine from APC Δ468 mice were stained with antibodies to CD11b (AlexaFluor 488), Gr1 (AlexaFluor 594) and DAPI. The merged images of CD11b with DAPI (a, CD11b green, DAPI gray), Gr1 with DAPI (b, Gr1 red, DAPI gray), and ProSense-680 with DAPI (c, ProSense-680 blue, DAPI gray) were produced with the “RGB gray” plug-in of Image J. The “colocalization finder” plug-in produced images where the colocalized pixels appear white while the ProSense-680 was red (d&e), the CD11b was green (d, colocalization analysis of ProSense-680 and CD11b staining) and the Gr1 was green (e, colocalization analysis of ProSense-680 and Gr1 staining). ×400 magnification. Arrows mark a CD11b + Gr1 ProSense 680 + cell. Representative FACS dot-plots of MNCs prepared from polyposis intestine and ex vivo stained with ProSense-680 followed by CD11b and Gr1 staining. The live MNCs were gated for ProSence-680 + cells, which were analyzed for CD11b + Gr1 + (f, MDSCs) and CD11b + F4/80 + (g, macrophages) cells. Cumulative results of 6 FACS experiments showing % of CD11b + Gr1 + ProSense-680 + and CD11b + F4/80 + ProSense 680 + among total infiltrating MNCs. Note that among the ProSense-680 + cells (11±0.69% of total MNCs) over 75% were either CD11b + Gr1 + (3.4±0.6% of total MNCs) or CD11b + F4/80 (5.0±0.34% of total MNCs).

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: In Vivo, Staining, Produced, Ex Vivo

Journal: PLoS ONE

Article Title: Live Imaging of Cysteine-Cathepsin Activity Reveals Dynamics of Focal Inflammation, Angiogenesis, and Polyp Growth

doi: 10.1371/journal.pone.0002916

Figure Lengend Snippet: (a) Non-linear regression analysis of polyp number and diameter, assuming Gaussian distribution; APC Δ468 Ctsb −/− (continued line, open squares), and APC Δ468 (dotted line, closed triangles). Note that Cathepsin B −/− mice had fewer and smaller polyps. (b) Frequencies of ProSense-680 active leukocytes amongst total MNCs prepared from the intestine of APC Δ468 (open bar, 6.7%±086%) or APC Δ468 Ctsb −/− (filled bar, 11%±0.69%, P = 0.0037; unpaired t test with Welsh correction). (c) Frequencies of ProSense-680 active CD11b + Gr1 + (mean 0.56%) or CD11b + F4/80 + (mean 4.46%) cells, from the intestines of APC Δ468 (open bar) and APC Δ468 Ctsb −/− mice (filled bars); P<0.001, n = 6, 2way ANOVA. Note that Cathepsin B deficiency predominantly impacted the abundance of CD11b + Gr1 + cells. (d) Attenuation of polyposis in anti-TNFα treated mice (solid line, open squares, n = 6), as compared to the APC Δ468 (dotted line & closed triangles). (e) Frequencies of CD11b + Gr1 + amongst total intestine live MNCs; APC Δ468 Ctsb −/− intestine (light gray bar, 0.56±0.15%, P<0.001), anti-TNFα treated (dark gray bar, 0.71±0.22%, P<0.001), untreated APC Δ468 (open bar, 4.2±0.093%), wt control intestine (black bar, 0.15±0.051%). (f) Frequencies of CD11b + F4/80 + in the APC Δ468 (5.03±0.78%), APC Δ468 Ctsb −/− intestine (5.36±0.92%), and anti-TNFα treated intestine (dark gray bar, 3.0±0.67%).

Article Snippet: The frequencies of ProSense-680 + , CD11b + F4/80 + cells remain unchanged (mean 5.36% to 5.03% respectively).

Techniques: Control