Journal: International journal of molecular sciences

Article Title: Inhibiting CD44-ICD Attenuates LPS-Induced Initiation of Hepatic Inflammation in Septic Mice.

doi: 10.3390/ijms25168907

Figure Lengend Snippet: Figure 1. Effects of inhibiting CD44-ICD on LPS-induced hepatic inflammation and histological change in mice. Twenty mice were divided into four groups. N group mice were given only saline; D group mice were given DAPT alone; L group mice were given LPS alone; LD group mice were given DAPT 30 min before LPS. After 6 h, (A) AST levels, (B) ALT levels in serum, and (C) liver histopathological changes (10×, scale bar—100 µm; magnified 20×, scale bar—50 µm) by H&E staining were determined. Data are expressed as mean ± SD (n = 5). * p < 0.05 compared with N and D groups. # p < 0.05 compared with the L group.

Article Snippet: After blocking, the blots were incubated with iNOS (1:1000; BD Bioscience, Franklin Lakes, NJ, USA, 610432), p-IKK α/β (1:1000; Genetex, Washington, DC, USA, GTX9039), p-IκB (1:1000; Genetex, GTX00967), nuclear NF-κB (1:500; Genetex, GTX102090), IL-1β (1:1000, Santacruz, Dallas, TX, USA, sc-127420), CD44 (1:500; proteintech, 15675-1-AP), nuclear CD44-ICD (1:500; Cosmobio, Tokyo, Japan, #KAL-K0604), GAPDH (1:5000; abcam, ab9485), β tubulin (1:1000; abcam, ab6046), and Lamin B1 (1:1000; abcam, ab16048) in 5% nonfat skim milk.

Techniques: Saline, Staining

Journal: International journal of molecular sciences

Article Title: Inhibiting CD44-ICD Attenuates LPS-Induced Initiation of Hepatic Inflammation in Septic Mice.

doi: 10.3390/ijms25168907

Figure Lengend Snippet: Figure 2. Effects of inhibiting CD44-ICD on pro-inflammatory mediators in LPS-induced hepatic inflammation in mice and cells. Twenty mice were divided into 4 groups. N group mice were given only saline; D group mice were given DAPT alone; L group mice were given LPS alone; LD group mice were given DAPT 30 min before LPS. (A) hepatic IL-1β, (B) NO, and (C) iNOS expressions were determined six hours after LPS. The Chang cells were divided into five groups. N group, cells were treated only DMSO; L group, cells received LPS (100 ng/mL); LD5 group, cells received 5 mM DAPT 1 h before LPS; LD10 group, received 10 mM DAPT 1 h before LPS; LD 20 group, cells were received 20 mM DAPT 1 h before LPS. (D) IL-1β levels in Chang cells were determined 6 h after LPS was given. Data are expressed as mean ± SD (n = 5). * p < 0.05 compared with N and D groups. # p < 0.05 compared with the L group.

Article Snippet: After blocking, the blots were incubated with iNOS (1:1000; BD Bioscience, Franklin Lakes, NJ, USA, 610432), p-IKK α/β (1:1000; Genetex, Washington, DC, USA, GTX9039), p-IκB (1:1000; Genetex, GTX00967), nuclear NF-κB (1:500; Genetex, GTX102090), IL-1β (1:1000, Santacruz, Dallas, TX, USA, sc-127420), CD44 (1:500; proteintech, 15675-1-AP), nuclear CD44-ICD (1:500; Cosmobio, Tokyo, Japan, #KAL-K0604), GAPDH (1:5000; abcam, ab9485), β tubulin (1:1000; abcam, ab6046), and Lamin B1 (1:1000; abcam, ab16048) in 5% nonfat skim milk.

Techniques: Saline

Journal: International journal of molecular sciences

Article Title: Inhibiting CD44-ICD Attenuates LPS-Induced Initiation of Hepatic Inflammation in Septic Mice.

doi: 10.3390/ijms25168907

Figure Lengend Snippet: Figure 3. Effects of inhibiting CD44-ICD on NF-κB signaling-related protein expressions in LPS- induced hepatic inflammation in mice. Twenty mice were divided into 4 groups. N group mice were given only saline; D group mice were given DAPT alone; L group mice were given LPS alone; LD group mice were given DAPT 30 min before LPS. (A) Western blotting analysis of p- IKK, (B) immunohistochemical staining analysis of p-IKK, (C) Western blotting analysis of p-IκB, (D) immunohistochemical staining analysis of p-IκB, and (E) Western blotting analysis of nuclear NF-κB expression in liver tissue was determined 6 h after LPS. The immunohistochemical positive expression were observed under a microscope (10× with a scale bar of 100 µm). Data are expressed as mean ± SD (n = 5). * p < 0.05 compared with N and D groups. # p < 0.05 compared with the L group.

Article Snippet: After blocking, the blots were incubated with iNOS (1:1000; BD Bioscience, Franklin Lakes, NJ, USA, 610432), p-IKK α/β (1:1000; Genetex, Washington, DC, USA, GTX9039), p-IκB (1:1000; Genetex, GTX00967), nuclear NF-κB (1:500; Genetex, GTX102090), IL-1β (1:1000, Santacruz, Dallas, TX, USA, sc-127420), CD44 (1:500; proteintech, 15675-1-AP), nuclear CD44-ICD (1:500; Cosmobio, Tokyo, Japan, #KAL-K0604), GAPDH (1:5000; abcam, ab9485), β tubulin (1:1000; abcam, ab6046), and Lamin B1 (1:1000; abcam, ab16048) in 5% nonfat skim milk.

Techniques: Saline, Western Blot, Immunohistochemical staining, Staining, Expressing, Microscopy

Journal: International journal of molecular sciences

Article Title: Inhibiting CD44-ICD Attenuates LPS-Induced Initiation of Hepatic Inflammation in Septic Mice.

doi: 10.3390/ijms25168907

Figure Lengend Snippet: Figure 4. Effects of inhibiting CD44-ICD on CD44 expression in LPS-induced hepatic inflammation in mice and cells. Twenty mice were divided into 4 groups. N group mice were given only saline; D group mice were given DAPT alone; L group mice were given LPS alone; LD group mice were given DAPT 30 min before LPS. (A) Western blotting analysis of hepatic CD44 expression and (B) immunofluorescence staining analysis of hepatic CD44 expression was determined 6 h after LPS. Positive immunofluorescence reaction for CD44 (red color) and DAPI nucleic acid staining (blue color) was observed (20×) under a microscope. The Chang cells were divided into five groups. N group, cells were treated with only DMSO; L group, cells received LPS (100 ng/mL); LD5 group, cells received 5 mM DAPT 1 h before LPS; LD10 group received 10 mM DAPT 1 h before LPS; LD 20 group, cells received 20 mM DAPT 1 h before LPS. (C) CD44 and (D) nuclear CD44-ICD expression in Chang cells were determined 6 h after LPS was given. Data are expressed as mean ± SD (n = 5). * p < 0.05 compared with N and D groups. # p < 0.05 compared with the L group.

Article Snippet: After blocking, the blots were incubated with iNOS (1:1000; BD Bioscience, Franklin Lakes, NJ, USA, 610432), p-IKK α/β (1:1000; Genetex, Washington, DC, USA, GTX9039), p-IκB (1:1000; Genetex, GTX00967), nuclear NF-κB (1:500; Genetex, GTX102090), IL-1β (1:1000, Santacruz, Dallas, TX, USA, sc-127420), CD44 (1:500; proteintech, 15675-1-AP), nuclear CD44-ICD (1:500; Cosmobio, Tokyo, Japan, #KAL-K0604), GAPDH (1:5000; abcam, ab9485), β tubulin (1:1000; abcam, ab6046), and Lamin B1 (1:1000; abcam, ab16048) in 5% nonfat skim milk.

Techniques: Expressing, Saline, Western Blot, Immunofluorescence, Staining, Microscopy

Journal: Stem Cell Research & Therapy

Article Title: Glioma-associated mesenchymal stem cells-mediated PD-L1 expression is attenuated by Ad5-Ki67/IL-15 in GBM treatment

doi: 10.1186/s13287-022-02968-z

Figure Lengend Snippet: Identification of MSCs derived from human glioma tissues. A and B Adherent growth patterns and fibroblastic morphology of GA-MSCs cultured in MSC media (× 100, scale bars = 1000 µm). C Immunofluorescence showed that GA-MSCs expressed MSC specific markers CD44 and CD105 (× 400, scale bars = 500 µm)

Article Snippet: The cells were incubated with primary antibodies against CD44 and CD105 (1:100, Proteintech, China), then overnight at 4 °C.

Techniques: Derivative Assay, Cell Culture, Immunofluorescence

Journal: Circulation Research

Article Title: Parallel Murine and Human Plaque Proteomics Reveals Pathways of Plaque Rupture

doi: 10.1161/CIRCRESAHA.120.317295

Figure Lengend Snippet: Protein interaction network analysis reveals numerous interactions of proteins that are differentially abundant in aortas of SR-uPA +/0 mice. A protein-protein relational network was built based on experimentally validated direct interactions. The network is comprised of 87 proteins, each portrayed as a circular node (all nodes are identified in Data Set V in the Data Supplement ). Key highly connected nodes (hubs) are labeled together with 2 members of the matrix metalloproteinase family of extracellular proteases and several extracellular matrix components. ACTB indicates beta actin; AGRN, agrin; BCAM, basal cell adhesion molecule; ELN, elastin; FBLN5, fibulin 5; FN1, fibronectin 1; HSPG2, heparan sulfate proteoglycan 2; LAMA5, laminin subunit alpha 5; LAMB2, laminin subunit beta 2; LAMC1, laminin subunit gamma 1; LTBP4, latent transforming growth factor binding protein 4; MMP2, matrix metalloproteinase 2; MMP3, matrix metalloproteinase 3; MYH9, myosin heavy chain 9; NID1, nidogen1; NID2, nidogen 2; and PLAU, urokinase-type plasminogen activator.

Article Snippet: HSPG2 was visualized with 0.375 μg/mL of HSPG2 antibody (PB9277; Boster Biological Technology) in 5% milk/PBST (phosphate-buffered saline/Tween) overnight.

Techniques: Labeling, Binding Assay