fas inhibitor c75  (Cayman Chemical)

Journal: mBio

Article Title: Antiviral Wolbachia strains associate with Aedes aegypti endoplasmic reticulum membranes and induce lipid droplet formation to restrict dengue virus replication

doi: 10.1128/mbio.02495-23

Figure Lengend Snippet: Disrupting Wolbachia -induced lipid droplets facilitates viral replication in w Mel- Aag2 cell line. Wolbachia -free and w Mel- Aag2 cell lines were mock-infected or DENV-2 infected at MOI 1 for 24 h. Prior to DENV-2 infection cells were pre-treated with C75 for 24 h, and C75 was maintained during the time of infection. ( A–B ) Cell-associated RNA and virus supernatant were collected from each cell line 24 h post-DENV-2-infection for analysis by qRT-PCR ( A ) and TCID 50 ( B ). Data are the median number of viral genome copies ± IQR relative to the RPS17 mosquito housekeeping gene ( A ) or the median TCID 50 /mL ± IQR ( B ). The numbers in parentheses represent the average number of Wolbachia per cell ( Wolbachia 16S rRNA/RPS17 ), determined in parallel wells for each independent experiment prior to DENV-2 inoculation. Data are representative of at least three ( A ) or two independent experiments ( B ) with triplicate biological replicates each. Statistical analyses were performed using Mann-Whitney test, where ** P < 0.001 and **** P < 0.0001. ( C ) Wolbachia -free (top panel) and w Mel- Aag2 cell lines (bottom panel) were stained with BODIPY (green) and DAPI (blue). Wolbachia was detected with an α-WSP antibody (red) and 4G2 hybridoma fluid against flavivirus group E antigen for DENV-2 staining (magenta). Slides were imaged as 3-dimensional z-stacks and 2D images generated by MIP using Fiji software. Scale bars = 15 µm. ( D ) LD numbers were quantified 24 h post-DENV-2 infection at MOI 1. LD numbers were analyzed using Fiji software. Data are the mean LD number ± SEM from at least 6 fields of view representing at least 80 cells in total, where each data point represents the mean LD number from a single field of view. Statistical analyses were performed by an unpaired two-tailed student’s t -test, where **** P < 0.0001. ( E ) TEM of Wolbachia -free and w Mel- Aag2 cell lines mock-infected or DENV-2 infected at MOI 1 for 24 h and treated with C75 as previously described. Scale bar = 500 nm. ER, endoplasmic reticulum; W, Wolbachia ; M, mitochondria; Vp, vesicle packets.

Article Snippet: Prior to DENV-2 infection cells were pre-treated with 1µM fatty acid synthase (FAS) inhibitor C75 (Abcam) or DMSO diluent only for 24 h. Cells were mock-infected or DENV-2-infected at MOI 1 for 24 h, and C75 was maintained during the time of infection in Aag2 cell culture medium with 10% FBS.

Techniques: Infection, Virus, Quantitative RT-PCR, MANN-WHITNEY, Staining, Generated, Software, Two Tailed Test

Journal: Stem Cells

Article Title: Increased lipogenesis is critical for self‐renewal and growth of breast cancer stem cells: Impact of omega‐3 fatty acids

doi: 10.1002/stem.3452

Figure Lengend Snippet: Characteristics of lipid metabolism in breast cancer stem cells. A,B, Representative images of mammosphere cells (MCF‐7‐CSC) derived from MCF‐7 cell line (A) and EMT‐CSC induced from HMLE‐Twist‐ER cells (B). C,D, Comparison of the profiles of SFA and MUFA (top and middle), and of C18:2n‐6 and C20:4n‐6 PUFA (bottom) in MCF‐7‐CSC (C) and EMT‐CSC (D) compared with their parental cells, respectively. E,F, Representative Western blot for protein expressions of FAS, SCD1, and FADS2 in MCF‐7‐CSC (E) and EMT‐CSC (F) compared with their parental cells, respectively. G,H, Relative densiometric bar graphs of FAS, SCD1, and FADS2 in MCF‐7‐CSC (G) and EMT‐CSC (H) compared with their parental cells, respectively. * P < .05; ** P < .01 vs parental cells. All the experiments were repeated at least three times. CSC, cancer stem cells; EMT, epithelial‐mesenchymal transition; MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids; SFA, saturated fatty acids

Article Snippet: FAS inhibitor (C75), SCD1 inhibitor (CAY‐10566), and D6D inhibitor (SC‐26196) were purchased from Cayman.

Techniques: Derivative Assay, Comparison, Western Blot

Journal: Stem Cells

Article Title: Increased lipogenesis is critical for self‐renewal and growth of breast cancer stem cells: Impact of omega‐3 fatty acids

doi: 10.1002/stem.3452

Figure Lengend Snippet: Increased expression of the lipogenic enzymes in human breast cancer tissues. A,C,E, Representative image of IHC analysis for SCD1 (A), FAS (C), and FADS1 (E) expressions in normal and tumor tissues. ANT, tumor‐adjacent normal tissue; T, tumor. B,D,F, Representative image of immunohistochemistry (IHC) analysis for SCD1 (B), FAS (D), and FADS1 (F) expressions between various clinical and tumor grades (G1‐3). *** P < .001 vs vehicle‐treated control cells. Alphabet letters were used to show the significant difference between groups. Each subfigure contains representative image (top) and quantitative results (bottom). The quantitative results were plotted by the values of minimum to maximum

Article Snippet: FAS inhibitor (C75), SCD1 inhibitor (CAY‐10566), and D6D inhibitor (SC‐26196) were purchased from Cayman.

Techniques: Expressing, Immunohistochemistry, Control

Journal: Stem Cells

Article Title: Increased lipogenesis is critical for self‐renewal and growth of breast cancer stem cells: Impact of omega‐3 fatty acids

doi: 10.1002/stem.3452

Figure Lengend Snippet: n‐3 PUFA inhibit tumor growth of breast cancer stem cells in xenograft mouse model. A, Representative image of tumor‐bearing mice. Prior to implanting 1.5 × 10 7 of MCF‐7‐CSC to nude mice, these mice were orally gavaged by either fish oil (n‐3 PUFA) or corn oil (control) for a month and continued giving oils until the end of the experiment. B, Tumor growth of the nude mice in each group. C, Relative tumor weight by body weight. D, The ratio of n‐6/n‐3 PUFA. E, The ratio of MUFA (C16:1 + C18:1)/SFA (C16:0 + C18:0). F, The ratio of C20:4/C18:2. G,H, Protein levels (G) and mRNA levels (H) of SCD1, FAS, FADS2, and COX2 between control and n‐3 PUFA groups. n = 3 to 4. * P < .05; ** P < .01; *** P < .001 vs control. MUFA, monounsaturated fatty acids; PUFA, polyunsaturated fatty acids; SFA, saturated fatty acids

Article Snippet: FAS inhibitor (C75), SCD1 inhibitor (CAY‐10566), and D6D inhibitor (SC‐26196) were purchased from Cayman.

Techniques: Control