Journal: bioRxiv

Article Title: Cachexia and fibrosis are costs of chronic IL-1R-mediated disease tolerance in T. gondii infection

doi: 10.1101/783316

Figure Lengend Snippet: A-C Western blot for alpha smooth muscle actin (α-SMA) or GAPDH on liver (A) , quadriceps (B) , and vWAT (C) tissue lysate at 9 wpi. Representative of at least 3 independent experiments. Each lane represents an individual mouse. D-F , Picrosirius red staining on formalin-fixed paraffin-embedded liver (D) , gastrocnemius (E) , or vWAT (F) using polarized light at 9 weeks post-infection. Representative images shown at left. Picrosirius red staining was quantified in ImageJ (right) and pixel density represented as % of each field of view. Each point represents a field of view (N=3 mice per group, 5-10 fields of view per mouse were quantified). Scale bars represent 50 μm. D-E are representative of at least 2 experiments and (F) is pooled between two independent experiments. Error bars are standard error of the mean. Statistical outliers were removed using the ROUT method (Q=1%), in D-F. *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired Student’s T test.

Article Snippet: The next morning, samples were washed 3 times in PBS/0.1% Triton-X and were incubated in secondary antibody (Novus NBP1-75607 donkey anti-goat Dylight 594, 1:250 and LifeTech A21206 donkey anti-rabbit AF488, 1:200), and the directly conjugated α-SMA primary (Novus Biologicals NBP2-34760APC, 1:400) for 1 hour at room temperature, diluted in PBS/0.1% Triton-X.

Techniques: Western Blot, Staining, Formalin-fixed Paraffin-Embedded, Infection

Journal: bioRxiv

Article Title: Cachexia and fibrosis are costs of chronic IL-1R-mediated disease tolerance in T. gondii infection

doi: 10.1101/783316

Figure Lengend Snippet: A-C Western blot for alpha smooth muscle actin (α-SMA) or GAPDH on liver (A), vWAT (B), and quad (C) tissue lysate at 2 or 9 wpi. Representative of at least 2 independent experiments. Each lane represents an individual mouse. D-F Picrosirius red staining on formalin-fixed paraffin-embedded liver (D), gastrocnemius (E), or vWAT (F) using brightfield (D) or polarized light (E-F) at 9 weeks post-infection. Picrosirius red staining was quantified in ImageJ (below) and pixel density represented as % of each field of view. N=3 mice per group, 5-10 fields of view per mouse were quantified Representative of 2 experiments. Error bars are standard error of the mean. Scale bars represent 50 μm. Statistical outliers were removed using the ROUT method (Q=1%), in E and F. *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001 by unpaired Student’s T test with Holm-Sidak method to correct for multiple comparisons.

Article Snippet: The next morning, samples were washed 3 times in PBS/0.1% Triton-X and were incubated in secondary antibody (Novus NBP1-75607 donkey anti-goat Dylight 594, 1:250 and LifeTech A21206 donkey anti-rabbit AF488, 1:200), and the directly conjugated α-SMA primary (Novus Biologicals NBP2-34760APC, 1:400) for 1 hour at room temperature, diluted in PBS/0.1% Triton-X.

Techniques: Western Blot, Staining, Formalin-fixed Paraffin-Embedded, Infection

Journal: bioRxiv

Article Title: Cachexia and fibrosis are costs of chronic IL-1R-mediated disease tolerance in T. gondii infection

doi: 10.1101/783316

Figure Lengend Snippet: A , Cytokines in liver lysates from mice 9 wpi were measured by ELISA. Data are presented as fold change relative to the mean of uninfected levels. N=4-13 mice per group, pooled from two independent experiments. B-C , Immunofluorescence labeling of nuclei (DAPI white) IL-1α (green), CD45 (red) and collagen1α1 (blue) ( B ) nuclei (DAPI white) α-smooth muscle actin (green), IL-1R (red) and collagen1α (blue) in the liver of uninfected or 9 wpi WT or IL-1R -/- mice ( C ). Inset, arrow head represents α-smooth muscle actin, IL-1R co-staining cells (arrow heads). ( B-C ) represent maximum intensity projections of 9-13 μm thick z-stacks. Scale bar represents 50 μm. Error bars are standard error of the mean. *P < 0.05; **P < 0.01; ***P < 0.001 by unpaired Student’s T test with Holm-Sidak method to correct for multiple comparisons.

Article Snippet: The next morning, samples were washed 3 times in PBS/0.1% Triton-X and were incubated in secondary antibody (Novus NBP1-75607 donkey anti-goat Dylight 594, 1:250 and LifeTech A21206 donkey anti-rabbit AF488, 1:200), and the directly conjugated α-SMA primary (Novus Biologicals NBP2-34760APC, 1:400) for 1 hour at room temperature, diluted in PBS/0.1% Triton-X.

Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Labeling, Staining

Journal: bioRxiv

Article Title: Cachexia and fibrosis are costs of chronic IL-1R-mediated disease tolerance in T. gondii infection

doi: 10.1101/783316

Figure Lengend Snippet: A-C , MEF cells were incubated with media, 10ng/mL IL-1α or 5 ng/mL TGFβ-1 for 48 hours. After fixation, MEFs were stained for F-actin, alpha-smooth muscle actin (α-SMA), phalloidin and nuclei and cell spreading was quantified in (B) , and levels of α-SMA expression were quantified in terms of pixels/cell (C). D-F , Primary hepatic stellate cells (HSCs) were isolated from uninfected mouse livers, and FACS sorted based on endogenous retinoid fluorescence. Retinoid fluorescence was validated by UV photobleaching ( D ). E-F , HSCs were seeded onto 4kPa hydrogels coated with 10ug/mL of fibronectin and cultured with 10ng/mL IL-1α, 10 ng/mL TGF-β, or media alone for 48hrs and then fixed and stained for F-actin and α-SMA and imaged by confocal microscopy. Scale bar represents 50 μm. Total cell area quantified in ( F ) Error bars are standard error of the mean. *P < 0.05; **P < 0.01; ***P < 0.001, ****P < 0.0001 by unpaired Student’s T test.

Article Snippet: The next morning, samples were washed 3 times in PBS/0.1% Triton-X and were incubated in secondary antibody (Novus NBP1-75607 donkey anti-goat Dylight 594, 1:250 and LifeTech A21206 donkey anti-rabbit AF488, 1:200), and the directly conjugated α-SMA primary (Novus Biologicals NBP2-34760APC, 1:400) for 1 hour at room temperature, diluted in PBS/0.1% Triton-X.

Techniques: Incubation, Staining, Expressing, Isolation, Fluorescence, Cell Culture, Confocal Microscopy

Journal: Journal of nanobiotechnology

Article Title: Evaluating the pro-survival potential of apoptotic bodies derived from 2D- and 3D- cultured adipose stem cells in ischaemic flaps.

doi: 10.1186/s12951-024-02533-1

Figure Lengend Snippet: Fig. 6 3D-ABs suppress oxidative stress and cell death, and stimulated angiogenesis in ischaemic flaps. (A) F-CHP staining for damaged collagen detec tion in the skin on POD7. Scale bars, 100 μm. (B) The three groups’ quantified F-CHP intensity (n = 6). (C) CD31 and α-SMA IF staining in the flap on POD7. Scale bars, 50 μm. (D) Quantified CD31/α-SMA-positive blood vessel density among the three groups (n = 6). (E) Dead cells in flap tissue sections on POD7 under the detection of TUNEL staining. Scale bar, 20 μm. (F) TUNEL-positive cell percentage in the dermal layer, quantified for each of the three groups (n = 6). (G) DHE staining of the skin tissues on POD7 among the three groups. Scale bars, 50 μm. (H) Quantified DHE among the three groups (n = 6). SEM error bars are used. Significance (*): p value < 0.05; equal variances ANOVA with LSD post hoc analysis or unequal variances Dunnett’s T3 technique

Article Snippet: The primary antibodies utilized were specific to CD31 (1:200), CD68 (1:200), α-SMA (1:200; Proteintech, 67735-1-Ig), iNOS (1:200), and Arg1 (1:200).

Techniques: Staining, TUNEL Assay

Journal: bioRxiv

Article Title: The Aging Microenvironment as a Determinant of Immune Exclusion and Metastatic Fate in Pancreatic Cancer

doi: 10.1101/2025.05.08.652966

Figure Lengend Snippet: A, Flow cytometry-based characterization of isolated murine CAFs for pan-CAF markers, including podoplanin (PDPN) and fibroblast-specific protein 1 (FSP1). B, Immunoblot showing the expression of fibroblast marker α-SMA in young and old CAFs, and the epithelial marker E-Cadherin in KPC cells. β-actin serves as a loading control. C−D , Table summarizes the incidence of metastases across various organ sites in old and young recipients, following heterochronic co-implantation of murine CAFs with KPC cells.

Article Snippet: Protein samples were separated by SDS-PAGE electrophoresis and then transferred to nitrocellulose membranes using Trans-Blot Turbo Transfer System (Bio-Rad), followed by incubation with primary antibodies: rabbit α-SMA (CST, 1:1000), rabbit E-cadherin (CST, 1:1000), and mouse β-Actin (Sigma, A2228, 1:5000).

Techniques: Flow Cytometry, Isolation, Western Blot, Expressing, Marker, Control

Journal: Redox Biology

Article Title: Chicoric acid prevents PDGF-BB-induced VSMC dedifferentiation, proliferation and migration by suppressing ROS/NFκB/mTOR/P70S6K signaling cascade

doi: 10.1016/j.redox.2017.11.012

Figure Lengend Snippet: CA abrogated PDGF-BB-induced VSMC dedifferentiation. VSMCs were pretreated with various concentrations (10, 50 and 100 μM) of CA for 6 h followed by stimulation with PDGF-BB (20 ng/mL) for 24 h. ( A ) Western blot was employed to quantitate the expression levels of contractile protein α-SMA, SMMHC, SM22α and synthetic proteins OPN. ( B ) Bar graph showing the relative protein level of α-SMA, SMMHC, SM22α and OPN. ( C ) Bar graph showing the relative mRNA level ofα-SMA, SMMHC, SM22α and OPN. Values are mean±SE. * P < 0.05 vs. Control, † P < 0.05 vs. PDGF-BB. n = 6 for each group.

Article Snippet: Antibodies against α-SMA, SM22α, PCNA, cyclin D1, P27 and horseradish peroxidase conjugated secondary antibodies were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Western Blot, Expressing, Control