Journal: Frontiers in cell and developmental biology

Article Title: Absence of Desmin Results in Impaired Adaptive Response to Mechanical Overloading of Skeletal Muscle.

doi: 10.3389/fcell.2021.662133

Figure Lengend Snippet: FIGURE 2 | Transition toward oxidative fibers is not impaired in DesKO plantaris in response to OVL. Representative images (A–D) of plantaris section immunostained for MHC-2a (red), MHC-2b (blue) and perlecan (green) from both genotype (Ctr and DesKO) in basal condition or after 1 month of OVL. Percentage of MHC-2a positive fibers (E) or MHC-2b positive fibers (F). Percentage of the surface of muscle section demonstrating succinate dehydrogenase activity (G). Scale bar = 100 µm. Data are given as means ± SEM. DesKO, Desmin knock-out mice; Ctr, Control mice; OVL, mechanical overloading; MHC, myosin heavy chain; SDH, succinate dehydrogenase. ns: non-significant, *p < 0.05, **p< 0.01, ***p< 0.001.

Article Snippet: Briefly, the sections were incubated with primary antibodies against heparan sulfate proteoglycan (Perlecan) (1:400, rat monoclonal, Millipore), myosin heavy chain (MHC)-2a (1:50, mouse monoclonal, clone SC-71, Developmental Studies Hybridoma Bank, University of Iowa) or MHC-2b (1:5, mouse monoclonal, clone BF-F3, Developmental Studies Hybridoma Bank, University of Iowa).

Techniques: Activity Assay, Knock-Out, Control

Journal: bioRxiv

Article Title: The level of endothelial glycocalyx maturity modulates interactions with charged nanomaterials

doi: 10.1101/2024.09.10.611831

Figure Lengend Snippet: (A) Representative confocal microscope image of endothelial cells exposed to AuNP+ (3 μg/mL; white) for 2 h and counterstained for the (A) plasma membrane (red; CellMask™), (B) syndecan-1 (green; antibody clone DL-101), and CD44 (green; antibody clone Hermes-1) and (C) HS (green; antibody clone 10E4), HA (green; HA binding protein) and glycans (WGA; red) and nuclei (blue; Hoechst-33342). Scale bar is 40 μm. Manders’ colocalization coefficient (MCC) analysis for the extent of AuNP+ (white pixels) colocalized with (A) cell membrane, (B) glycocalyx proteins or (C) glycans. Data are mean ± SD (n=9).

Article Snippet: The cells were then washed three times in PBS and blocked with goat serum (1% v/v) in PBST (PBS with Tween-20 (0.05% w/v); blocking solution) at room temperature (RT) for 1 h. Cells were then incubated with anti-heparan sulfate (HS) (2 μg/mL, clone 10E4, Amsbio), anti-syndecan-1 (2 μg/mL, clone DL-101, Santa Cruz Biotechnology), anti-CD44 (2 μg/mL, clone Hermes-1, Developmental Studies Hybridoma Bank), or anti-VE-cadherin (1 μg/mL, Abcam) antibodies or biotinylated hyaluronan (HA) binding protein (5 μg/ml, Sigma-Aldrich) diluted in the blocking solution for 16 h at 4 °C then washed twice with PBST.

Techniques: Microscopy, Membrane, Binding Assay

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: 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: Discover oncology

Article Title: Preclinical efficacy of CBR-5884 against epithelial ovarian cancer cells by targeting the serine synthesis pathway.

doi: 10.1007/s12672-024-01013-0

Figure Lengend Snippet: Fig. 5 CBR-5884-impaired stemness of EOC cells and enhanced chemotherapy sensitivity. A SKOV3 and ID8 cells were treated with CBR- 5884 (50 μM) or dimethyl sulfoxide (DMSO; control) for 5 days, and tumorsphere formation ability was accessed by sphere formation assay. Representative images (left) and quantification data (right) are shown. B Cell cytometry was performed to detect CD44 expression in SKOV3 and ID8 cells after treatment with 30 μM CBR-5884 for 48 h. Representative images (left) and quantification data (right) are shown. C Quanti- tative polymerase chain reaction assay was conducted to measure the mRNA expression levels of stem-related genes in SKOV3 cells, includ- ing ALDH1A1, NANOG, SOX2, EPCAM, and POU5F1 after treatment with 30 μM CBR-5884 for 48 h. D SKOV3 and ID8 cells were treated with DMSO, carboplatin (20 μM), CBR-5884 (30 μM), and carboplatin (20 μM) plus CBR-5884 (30 μM) for 72 h. Cell viability was measured using CCK-8 assay

Article Snippet: To detect CD44-positive cells, a FITC anti-mouse CD44 (IM7; 1:200 dilution, Cat. No. FITC-65117; Proteintech, USA) antibody was used.

Techniques: Control, Tube Formation Assay, Cytometry, Expressing, Polymerase Chain Reaction, CCK-8 Assay