Journal: Chembiochem : a European journal of chemical biology

Article Title: Hypoxia Modulates Cellular Endocytic Pathways and Organelles with Enhanced Cell Migration and 3D Cell Invasion.

doi: 10.1002/cbic.202300506

Figure Lengend Snippet: Figure 4. CD98 immunostaining under hypoxia: Hypoxia was induced in the MDA-MB-231 cells. (A) After the hypoxia induction and galectin3 uptake, CD98 immunostaining was performed. The scale bar represents 20 μm. (B) Quantification of fluorescence intensity of CD98. (C) Quantification of fluorescence intensity of galectin3. Histobars represent the meanSD of normalized fluorescence intensity of 30 cells. Asterisks denote significant difference from the control, with *p0.05, **p0.01, ***p0.001, ****p0.0001 and ns represents non-significant difference from control.

Article Snippet: Mouse monoclonal antibodies to human CD98 were purchased from BioRad.

Techniques: Immunostaining, Fluorescence, Control

Journal: bioRxiv

Article Title: Wnts are endothelial cell-derived PKD1/PKD2-dependent autocrine/paracrine vasodilators

doi: 10.64898/2026.03.17.712518

Figure Lengend Snippet: (A) Representative RT-PCR showing that mRNA for PKD1 is present in isolated endothelial cells from tamoxifen-treated Pkd1 fl/fl but absent in isolated endothelial cells from tamoxifen-treated Pkd1 fl/fl :Cdh5(PAC)-CreERT2 mice. Representative of 5 independent experiments for each genotype. (B) Representative Western blots illustrating PKD1, PKD2, AT1, eNOS, Fzd-7 and actin proteins in mesenteric arteries of Pkd1 fl/fl and Pkd1 ecKO mice. (C) Mean data from experiments shown in panel B. Significance was assessed using Student t-tests, n=5 independent mesenteric arterial lysates for each genotype and each protein. (D) Immunofluorescence images (representative of three mesenteric arteries from three mice for each genotype) illustrating that PKD1 (Alexa Fluor 546) is abolished in endothelial cells of en face mesenteric arteries from Pkd1 ecKO mice. CD31 (Alexa Fluor 488) and DAPI are also shown. Scale bars, 50 μm. (E) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular Wnt9b (3 μg/ml) applied in continuous flow in pressurized (80 mmHg) mesenteric arteries from Pkd1 fl/fl and Pkd1 ecKO mice. (F) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular boiled Wnt9b (3 µg/ml) applied under continuous flow (10 dyn/cm 2 ) in pressurized (80 mmHg) mesenteric arteries from Pkd1 fl/fl and Pkd1 ecKO mice. (G) Mean data from experiments shown in panels E and F, and the modulation of flow and Wnt9b-mediated vasodilation by SRI37892 (Fzd-7 Inh, 5 µM). Significance was assessed using a two-way ANOVA with Holm-Sidak post hoc multiple comparisons test. n=10 arteries from 7 mice of each genotype for flow, flow+ Wnt9b and Wnt9b. n=7 arteries from 5 mice for Fzd-7 Inh+flow and Fzd-7 Inh+flow+Wnt9b. n=5 arteries from 5 mice for each genotype for flow + boiled Wnt9b. (H) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular Wnt5a (3 µg/ml). (I) Diameter responses to intravascular flow (10 dyn/cm 2 ) and intravascular boiled Wnt5a (3 µg/ml). (J) Mean data from experiments shown in panels H and I and the modulation of flow and Wnt5a-induced vasodilation by SRI37892 (Fzd-7 Inh, 5 μM). Significance was assessed using two-way ANOVA with Holm-Sidak post hoc multiple comparisons test. n=8 arteries from 6 mice from Pkd1 fl/fl for flow, flow+ Wnt5a. n=5 arteries from 5 mice from Pkd1 ecKO for flow, flow+ Wnt5a. n=5 arteries from 5 mice of each genotype for flow + boiled Wnt5a. n=8 arteries from 6 mice from Pkd1 fl/fl for flow+ Fzd-7 Inh and flow+ Wnt5a+ Fzd-7 Inh.

Article Snippet: Membranes were blocked with 5% milk or 5% BSA and incubated with one of the following primary antibodies: PKD1 (Polycystic Kidney Disease Research Resource Consortium, Baltimore), PKD2 (Alomone), eNOS (Abcam), p-eNOS (Cell Signaling), Wnt9b (R&D Systems), Wnt5a (R&D Systems), AT1 receptor (Alomone) or actin (Cell Signaling) overnight at 4°C.

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Western Blot, Immunofluorescence

Journal: Cells

Article Title: Echinococcus granulosus -Induced Liver Damage Through Ferroptosis in Rat Model

doi: 10.3390/cells14050328

Figure Lengend Snippet: (A) Ultra-structure of hepatocytes near cysts at different stages of infection with PSCs (0, 1, 3, and 6 months); mitochondria are indicated by white arrows,PSC are indicated by black arrows( n = 3 rats/group, scale bars represent 2 μm and 1 μm). ( B ) Fe 2+ , ( C ) GSH, ( D ( a , b )) ROS, ( E ) MDA, ( F ) SOD, and ( G ) LDH of hepatocytes near cysts at different stages of infection with PSCs. ( H ( a , b )) Western blotting of protein expression levels of TFRC, GPX4, FTH1, NOX1, SLC3A2, and SLC7A11 in ferroptosis signaling pathway in rat liver tissue at different stages of infection with PSCs ( n = 3 rats/group)); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (Student t test).

Article Snippet: The antibodies used in this study were as follows: TFRC (Boster, 1: 500, Rabbit, BA0462-2, Wuhan, China); Caspase-3 (Boster, 1:500, Rabbit, BA3257, Wuhan, China); GSDMD (Proteintech Group, 1:800, Rabbit, 20770-1-AP, Wuhan, China); LC3I/II (Abcam, 1:1000, Rabbit, ab192890, Shanghai, China); Anti-GPX4 Antibody (Boster, 1:400, Rabbit, BM5231,Wuhan, China); Anti-FTH1 Antibody (Boster, China, 1:400, Rabbit, BM4487,Wuhan, China); Anti-NOX1 Antibody (Boster, China, 1:400, Rabbit, BA3720,Wuhan, China); Anti-CD98 Antibody (SLC3A2, Boster, China, 1:400, Rabbit, A01794-1,Wuhan,China); Anti-xCT Antibody (SLC7A11,Abcam,1:400, Rabbit, A01794-1, Shanghai, China); β-actin (Sino Biological, 1:1000, Mouse, 100166-MM10, Beijing, China).

Techniques: Infection, Western Blot, Expressing

Journal: Cells

Article Title: Echinococcus granulosus -Induced Liver Damage Through Ferroptosis in Rat Model

doi: 10.3390/cells14050328

Figure Lengend Snippet: ( A ) Fe 2+ in normal BRL cells, PSCs and BRL co-cultured cells, and PSCs +Ferrostatin-1 and BRL co-cultured cells. ( B ) GSH, ( C ) GSSH, ( D ) GSH/GSSH, ( E ) SOD, ( F ) LDH, and ( G ) MDA concentration detection ( n = 6 rats/group). ( H ( a , b )) Western-blotting detection of expression levels of cell ferroptosis signaling pathway proteins in each group, such as TFRC, GPX4, FTH1, NOX1, SLC3A2, andSLC7A11 ( n = 3 rats/group); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 (Student t test).

Article Snippet: The antibodies used in this study were as follows: TFRC (Boster, 1: 500, Rabbit, BA0462-2, Wuhan, China); Caspase-3 (Boster, 1:500, Rabbit, BA3257, Wuhan, China); GSDMD (Proteintech Group, 1:800, Rabbit, 20770-1-AP, Wuhan, China); LC3I/II (Abcam, 1:1000, Rabbit, ab192890, Shanghai, China); Anti-GPX4 Antibody (Boster, 1:400, Rabbit, BM5231,Wuhan, China); Anti-FTH1 Antibody (Boster, China, 1:400, Rabbit, BM4487,Wuhan, China); Anti-NOX1 Antibody (Boster, China, 1:400, Rabbit, BA3720,Wuhan, China); Anti-CD98 Antibody (SLC3A2, Boster, China, 1:400, Rabbit, A01794-1,Wuhan,China); Anti-xCT Antibody (SLC7A11,Abcam,1:400, Rabbit, A01794-1, Shanghai, China); β-actin (Sino Biological, 1:1000, Mouse, 100166-MM10, Beijing, China).

Techniques: Cell Culture, Concentration Assay, Western Blot, Expressing

Journal: STAR Protocols

Article Title: A flow-cytometry-based protocol using diverse cell types for detecting autoantibodies from human plasma and serum samples

doi: 10.1016/j.xpro.2021.100924

Figure Lengend Snippet: Sample plating layout To aid in the differentiation of control reactions from sample reactions, designate a row within a plate to controls (top row), followed by samples. Cells within the control wells will not be used to screen biological samples. The IgG Positive control will receive both the anti-CD98 antibody, as well as the staining cocktail. Single color controls, in concert with the unstained control will provide preliminary guidance as to where to draw positive-event gates. The placements of these gates are later refined by the boundaries set by the fluorescence-minus-one controls. We separated healthy human control samples from biological human samples of interest. These healthy human controls can further refine the lower limits of positive-event gates, as it is hypothesized that healthy human controls lack or have minimal circulating autoantibodies.

Article Snippet: Human anti-CD98 (20 μg/mL for 5 × 10 4 cells) , Absolute Antibody , Ab00361–10.0.

Techniques: Positive Control, Staining, Fluorescence

Journal: STAR Protocols

Article Title: A flow-cytometry-based protocol using diverse cell types for detecting autoantibodies from human plasma and serum samples

doi: 10.1016/j.xpro.2021.100924

Figure Lengend Snippet:

Article Snippet: Human anti-CD98 (20 μg/mL for 5 × 10 4 cells) , Absolute Antibody , Ab00361–10.0.

Techniques: Recombinant, Electron Microscopy, Flow Cytometry