Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: cPLA2 and desaturases underlie the tau hyperphosphorylation offset induced by BACE knock-down in neuronal primary cultures.

doi: 10.1016/j.bbadis.2018.08.028

Figure Lengend Snippet: Fig. 3. SCD1 silencing increased AT8 and COX2 levels after BACE1 silencing in glutamate-mediated excitotoxicity. Primary cortex neurons were doubly transduced with SCR, BACE1 shRNA-miR and cPLA2 shRNA and treated with 125 μM glutamate for 20 min. After 24 h, (A) SCD1, (B) tAT8, (C) COX2 and (D) INOS protein levels were detected by Western blotting. Actin and β-III tubulin were used as loading controls. Relative Units = RU, n = 4, and each experiment was performed in duplicate. All values were normalized to the control neurons. ANOVA with Tukey's test was used for comparisons. * Comparison between each treatment in the same group. # Comparison between shSCRmiR and shcPLA2miR groups. *,#p < 0.05; **,##p < 0.01; ***,##p < 0.001.

Article Snippet: To increase the expression of the SCD1 (mouse activation plasmid; Santa Cruz Biotechnology, Santa Cruz, CA, USA); and FADS6 (mouse activation plasmid; Santa Cruz Biotechnology, Santa Cruz, CA, USA) genes in the neuronal primary cultures, we used the CRISPR activation plasmid (m) as a system for SCD1 and FADS6 transcription activation as a synergistic activation mediator (SAM).

Techniques: Transduction, shRNA, Western Blot, Control, Comparison

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: cPLA2 and desaturases underlie the tau hyperphosphorylation offset induced by BACE knock-down in neuronal primary cultures.

doi: 10.1016/j.bbadis.2018.08.028

Figure Lengend Snippet: Fig. 5. FADS6 silencing increased AT8 and COX2 levels after BACE1 silencing in glutamate-mediated excitotoxicity. Primary cortex neurons were doubly transduced with SCR, BACE1 shRNA-miR and FADS6 shRNA and treated with 125 μM glutamate for 20 min. After 24 h, the proteins were collected. In (A) the SCD1, (B) the AT8, (C) COX2 and (D) the INOS protein levels were detected by Western blotting. Actin and β-III tubulin were used as the loading controls. Relative Units = RU, n = 4 and each experiment was performed in duplicate. All values were normalized to the control neurons. ANOVA with Tukey's test was used for comparisons. * Comparison between each treatment in the same group. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: To increase the expression of the SCD1 (mouse activation plasmid; Santa Cruz Biotechnology, Santa Cruz, CA, USA); and FADS6 (mouse activation plasmid; Santa Cruz Biotechnology, Santa Cruz, CA, USA) genes in the neuronal primary cultures, we used the CRISPR activation plasmid (m) as a system for SCD1 and FADS6 transcription activation as a synergistic activation mediator (SAM).

Techniques: Transduction, shRNA, Western Blot, Control, Comparison

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Glavonoid-rich oil supplementation reduces stearoyl-coenzyme A desaturase 1 expression and improves systemic metabolism in diabetic, obese KK-A y mice.

doi: 10.1016/j.biopha.2021.111714

Figure Lengend Snippet: Fig. 9. GRO supplementation reduced the protein levels of SCD1 and PPARα and increased the level of phosphorylated AMPK in the liver. (A) Western blot analysis of SCD1 in the liver of 20-week-old mice. (B) Western blot analysis of PPARα in the liver of 20-week-old mice. (C) The ratio of phosphorylated AMPK to total AMPK levels in the liver of 20-week-old mice, as determined by Western blot analysis. Cont: control mice; GRO8: 0.8% GRO mice. The data were analyzed with Student’s t- test. *p < 0.05, *p < 0.01. Values are means ± SD (n = 5).

Article Snippet: After electrophoresis, the separated proteins were transferred to a polyvinylidene difluoride membrane (Immobilon, 0.2 μm pore, (Merck Millipore Burlington, MA, USA)) and incubated overnight at 4 ◦C with the following primary antibodies: polyclonal rabbit anti-mouse PPARα (1:3000, GeneTex, Los Angeles, CA, USA), β-actin (1:3000, GeneTex), total AMPKα1 (1:3000, GeneTex), phosphorylated AMPKα1 (Thr172) (1:1000, Cell Signaling Technology, Danvers, MA, USA), and SCD1 (1:1000, R&D Systems Minneapolis, MN, USA).

Techniques: Western Blot, Control

Journal: EMBO Reports

Article Title: The anti‐apoptotic Bcl‐2 protein regulates hair follicle stem cell function

doi: 10.15252/embr.202052301

Figure Lengend Snippet: A, B Western blot analysis for Bcl‐2 protein levels in K15ΔNLef1 and wild‐type epidermis and K15ΔNLef1 tumours ( n = 3 biological replicates) (A) and Bcl‐2 levels in tumours of K15ΔNLef1 and K15ΔNLef1;Bcl‐2 EOE mice ( n = 4 biological replicates) (B). C Immunofluorescence staining of K15ΔNLef1 and K15ΔNLef1;Bcl‐2 EOE tumours, stained for Krt14 (green) and the sebaceous differentiation marker Scd1 (red). Scale bar, 100 µm. D Western blot analysis for Scd1 and Krt14 protein levels in tumours of K15ΔNLef1 and K15ΔNLef1;Bcl‐2 EOE mice ( n = 6 biological replicates). E Immunofluorescence staining of Krt14 (red) and BrdU (green) in K15ΔNLef1 and K15ΔNLef1;Bcl‐2 EOE tumours and quantification of the proliferation rate in K14 + cells ( n = 3 biological replicates; mean ± standard error of mean (SEM); ns, not significant, two‐tailed unpaired Student's t ‐test). F Immunofluorescence staining of aCas3 (red) and Krt14 (green) in K15ΔNLef1 and K15ΔNLef1;Bcl‐2 EOE tumours and quantification of the apoptosis rate in K14 + cells ( n = 3 biological replicates; mean ± standard error of mean (SEM); ns, not significant, two‐tailed unpaired Student's t ‐test).

Article Snippet: The following primary antibodies were used: K15 (mouse, Neomarkers, 1:1,500), K14 (rabbit, Covance, 1:3,000), K10 (rabbit, Covance, 1:500), K6 (rabbit, Covance, 1:3,000), active Caspase 3 (rabbit, R&D Systems, 1:500), BrdU (mouse, BD Bioscience, 1:50), P‐Cadherin (rat, Invitrogen, 1:400), Nfatc1 (7A6, mouse, DSHB), Sox9 (rabbit, Millipore, 1:1,000), Scd1 (rat, R&D, 1:100), BrdU (rat, Oxford Biotechnology, 1:500), K15 (rabbit, Progen, 1:1,500), Integrin α6 (rat, BD, 1:1,000), γH2AX (rabbit, Cell Signaling, 1:200), K71, K72, K75, K85 (guinea pig, Progen, ready‐to‐use kit), Bcl‐2 (mouse, BD Bioscience, 1:200), Trp2 (goat, Santa Cruz, 1:200) and GFP‐AlexaFITC (goat, Rockland, 1:500).

Techniques: Western Blot, Immunofluorescence, Staining, Marker, Two Tailed Test