Journal: Food Science & Nutrition

Article Title: Dihydroquercetin Attenuates Silica‐Induced Pulmonary Fibrosis by Modulating the Gut Microbiota and the Serum Metabolites in Mice

doi: 10.1002/fsn3.71389

Figure Lengend Snippet: DHQ treatment attenuated silica‐induced pulmonary fibrosis in C57/BL6 mice. (A, B) DHQ treatment increased the body weight and decreased pulmonary index in silicosis model mice. The changes of body weight (C) The levels of pro‐inflammatory cytokines (IL‐1β, TNF‐α, and TGF‐β) in serum from different groups at day 21 were detected by ELISA assay. (D) Representative pictures (×200) of HE‐stained and Masson‐stained lung sections from mice on day 21 were shown. Bar = 100 μm. (E, F) The inflammation and fibrosis score numbers of 0–3, corresponding to the grades of –, +, ++, and +++, were evaluated by experienced pathologists in a blinded fashion. (G) Representative results of western blot for α‐SMA, collagen I and fibronectin in lung tissues and the quantification of results. Data are shown as mean ± SD. All experiments were repeated three times. # p < 0.05, ## p < 0.01 vs. the control group; * p < 0.05, ** p < 0.01 vs. the SiO 2 group.

Article Snippet: Sigma‐Aldrich provided the SiO 2 (Cat#S5631) particles (around 80% diameter 1‐5 μm), which were filtered through sedimentation following Stokes' law, underwent acidic hydrolysis, and were baked overnight at 200°C for 16 h. Interleukin‐1β (IL‐1β) (CSB‐E08054m), tumor necrosis factor‐α (TNF‐α) (CSB‐E04741m), and transforming growth factor‐β (TGF‐β) (CSB‐E04726m) were acquired from Cusabio Biotechnology in Wuhan, China.

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Control

Journal: International heart journal

Article Title: Ryanodine Receptor Type 2 Plays a Role in the Development of Cardiac Fibrosis under Mechanical Stretch Through TGFβ-1.

doi: 10.1536/ihj.16-572

Figure Lengend Snippet: Figure 2. Reduction of TGFβ1 expression by downregulation of RyR2in cardiomyocytes under mechani- cal stretch. A: Measurement of RyR2 mRNA level in cultured cardiomyocytes by Real-Time PCR. B: Expres- sion of RyR2 protein level in cardiomyocytes with Western Blotting. C: TGFβ1 in supernates of cultured cardiomyocytes was detected with ELISA analysis. D: Expression of TGFβ1 protein level in cardiomyocytes with Western Blotting. E: Measurement of tgfb1 mRNA level in cultured cardiomyocytes using Real-Time PCR. Values are expressed as mean ± SEM obtained from 3 independent experiments. shRyR2 indicates RyR2 knockdown lentiviral particle-infected cardiomyocytes; MS, cardiomyocytes under mechanical stretch; MS + ryr2 KD, RyR2 knockdown cardiomyocytes under mechanical stretch. *P < 0.05 versus control.

Article Snippet: ELISA assay: Supernates of cultured cardiomyocytes were collected and examined by Rat Transforming Growth Factor β1 ELISA Kit (CUSABIO BIOTECH, China) following the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Knockdown, Infection, Control

Journal: Frontiers in Immunology

Article Title: Megakaryocyte NLRP3 hyperactivation induces mild anemia and potentiates inflammatory response in mice

doi: 10.3389/fimmu.2023.1226196

Figure Lengend Snippet: Nlrp3 A350V/+ / Gp1ba- Cre KI/+ mice have elevated levels of caspase-1 and anemia. (A) Increased levels of both pro-caspase-1 and active enzyme in platelets from Nlrp3 A350V/+ / Gp1ba- Cre KI/+ mice (Mut) compared to control (Con) platelets. Representative blots out of two, each containing three different control and three mutant mice. (B, C) Densitometry of total (B) and cleaved/active caspase-1 (C) , normalized to α-tubulin. (D) Plasma levels of IL-1β in Nlrp3 A350V/+ / Gp1ba- Cre KI/+ vs. control mice; n = 7–8. (E) RBC counts and (F) hemoglobin contents in control vs. mutant mice, n = 7–19. (G) Representative femur bones from Nlrp3 A350V/+ / Gp1ba- Cre KI/+ mice vs. control mice. Note the reduced red color in the bones of mutant animals. (H) Hematoxylin–eosin staining of femur bones of control (upper panel) vs. mutant (lower panel) mouse. Note the lower numbers of nucleated cells in the mutant BM. Representative images out of 3 mice per group. (I) Strongly reduced Ter119+ (erythroid lineage) cells in the BM of mutant mice. (J) GATA-1 levels (MFI) in erythroid (Ter119+) cells in the BM of mutant vs. control mice, n = 5 for each group. (K) Fold change of the TGFβ content in MKs from the BM of control vs. Nlrp3 A350V/+ / Gp1ba- Cre KI/+ mice, n = 12. (L) Spleen weight normalized to the body weight, n = 7–14. (M) Staining of spleen sections of control (upper panel) vs. Nlrp3 A350V/+ / Gp1b α−Cre +/KI (lower panel) mice for Ter119 (pan-erythroid marker, green), CD3e (T-cell marker, pink), and CD19 (B-cell marker, blue). Representative sections out of three mice per group; ns, not significant.

Article Snippet: For intracellular staining, cells were permeabilized using PBS–Triton X-100 (0.1%, Abcam) for 10 min, before staining for GATA-1, anti-TGF-β1-APC (TW7-16B4, BioLegend), and FAM-FLICA Caspase-1 assay kit (Immunochemistry Technologies).

Techniques: Mutagenesis, Staining, Marker