Journal: Frontiers in Microbiology

Article Title: The CD147 Protein Complex Is Involved in Entry of Chikungunya Virus and Related Alphaviruses in Human Cells

doi: 10.3389/fmicb.2021.615165

Figure Lengend Snippet: CHIKV envelope association with the CD147 complex (A) Scheme of the used AP-MS approach. (B) CHIKV envelope Interaction partners of interest are listed. Examination of their plasma membrane presence (PM), presence of a transmembrane domain (TM), and MiST score (MiST) are indicated. Components of the CD147 protein complex retrieved in the CHIKV envelope affinity purification are indicated. (C–F) Western Blot of affinity purification of Strep-tagged Envelope protein. Detection of E2 and the interactors CD147, CD98, and SLC1A5. Affinity purification of untransfected cells (empty) were included as controls. On each blot input lysates of untransfected cells (empty) or E2 expressing cells (E2) and affinity purifications (AP) of the empty or E2 cells were loaded. For the CD147, CD98 and SLC1A5 blots 0.4 μl of input lysates and 10 μl of APs was loaded. For the E2 blot 5 μl of input lysates and 10 μl of APs was loaded.

Article Snippet: Membranes were washed with PBS-T and incubated with primary antibody anti-CD147 (ab232967, Abcam), anti-ASCT2 (SLC1A5) (V501, Cell Signaling), anti-CD98 (12206-T62, sino biologicals), anti-E1, anti-Strep (ab184224 or ab180957, Abcam) and anti-E2 (NR44002, Bei Resources) overnight at 4°C.

Techniques: Membrane, Affinity Purification, Western Blot, Expressing

Journal: Frontiers in Oncology

Article Title: Decrease of Intracellular Glutamine by STF-62247 Results in the Accumulation of Lipid Droplets in von Hippel-Lindau Deficient Cells

doi: 10.3389/fonc.2022.841054

Figure Lengend Snippet: Effects of STF-62247 on glutamine transporters. (A, B) mRNA relative expression, measured by RT-qPCR, of glutamine transporters SLC1A5, SLC7A5 and SLC3A2 influenced by (A) VHL status and (B) STF-62247. RCC4 VHL- and VHL+ were treated with 1.25 µM STF-62447 for 24 hr. Results for treated cells are compared to control cells (represented by the dotted line at 1). Results are presented as means and SEM of three independent experiments. (C) Glutamine transporters, SLC1A5, SL7A5 and SLC3A2, protein levels in RCC4 VHL- and VHL+ treated to STF-62247 for 24 and 48 hr. (D) Intracellular leucine was measured by LC-MS in RCC4 VHL- and VHL+ treated with 1.25 µM STF-62247 for 24 hr (N=6). Student’s t -tests were performed to compare results between VHL- and VHL+ cells (A) or between controls and treated cells (B, D) (*p < 0.05, **p < 0.01, ***p < 0.001). (E) Cell viability was evaluated by XTT assay in RCC4 VHL- and VHL+ cells. SLC1A5 and SLC7A5 inhibitors, GPNA and BCH respectively, were tested alone (concentrations from 0 to 1 mM) or combined with 1.25 µM STF-62247 (N=3). Two-Way ANOVA followed by Tukey’s test was performed to assess statistically significant results. Comparison between CTL and STF-62247 conditions for each concentration of inhibitor (GPNA or BCH) are denoted with the following statistical marks *p < 0.05, **p < 0.01 or ***p < 0.001. Comparison of each inhibitor concentrations (x-axis) are made with their respective control (0 mM columns, with or without STF-62247) and statistical significances are denoted by # p < 0.05, ## p < 0.01 or ### p < 0.001.

Article Snippet: Membranes were blocked using 5% skim milk diluted in a solution of 0.075% PBS-Tween (PBS-T) and incubated overnight in 3% BSA with specific primary antibodies against VHL, HIF-1α and HIF-2α (Cell signaling #68547, 14179, 59973), SLC1A5 and β-actin (Santa Cruz Biotechnologies sc-99002 and sc-47778), SLC7A5 (Medical & Biological Laboratories #BMP011), SLC3A2 (Aviva Systems Biology #OAAB00158), ASNS (Signalway #32909), SCD1 (Applied Biological Materials #ABM-G076), PLIN2 (Proteintech #15294-1-AP) and CPT1A (Abcam #ab220789).

Techniques: Expressing, Quantitative RT-PCR, Control, Liquid Chromatography with Mass Spectroscopy, XTT Assay, Comparison, Concentration Assay

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