Journal: Nutrients

Article Title: Vitamin C: A Novel Regulator of Neutrophil Extracellular Trap Formation

doi: 10.3390/nu5083131

Figure Lengend Snippet: Autophagy signaling is induced in Vitamin C deficient neutrophils. ( A ) Real time QPCR for ATG3, ATG5, ATG6, ATG7, and ATG8 mRNA from peritoneal PMNs of VitC sufficient and deficient Gulo −/− mice, ( N = 6 for each group, * p < 0.05). ( B ) Representative Western blot for expression of LC3B-I and LC3B-II from peritoneal PMNs of VitC sufficient and deficient Gulo −/− mice. Densitometry of LC3B-II/actin from peritoneal PMNs of VitC sufficient and deficient Gulo −/− mice ( N = 6 for each group, * p < 0.05). ( C ) Representative Western blot for expression of p62 and actin from peritoneal PMNs of VitC sufficient and deficient Gulo −/− mice. Densitometry of normalized p62 expression from peritoneal PMNs of VitC sufficient and deficient Gulo −/− mice ( N = 6 for each group, ns p = 0.3).

Article Snippet: Purified rabbit polyclonal antibodies to LC3B (L7543, Sigma-Aldrich), cleaved caspase-3 (#9661, Cell Signaling), caspase-3 (#9662, Cell Signaling), p62/SQSTM1 (NBP1-48320, Novus Biologicals), NFκB p65 (sc-109, Santa Cruz Biotechnology), Lamin B (sc-6216, Santa Cruz Biotechnology), and actin (sc-1616, Santa Cruz Biotechnology) were used in this study.

Techniques: Western Blot, Expressing

Journal: bioRxiv

Article Title: Inhibition of mTOR during a postnatal critical sensitive window rescues deficits in GABAergic PV cell connectivity and social behavior caused by loss of TSC1

doi: 10.1101/2020.03.29.014563

Figure Lengend Snippet: Western blot representative bands of LC3-I, LC3-II, p62 proteins ( A ) and pAMPK ( B ) and their quantification reveal that LC3-II and pAMPK expression are higher ( C , E ) in p14 Tg( Nkx2.1-Cre ); Tsc1 flox/flox vs Tsc1 Ctrl mice (Unpaired t-test: LC3-II *p=0.0314; pAMPK *p=0.0289), while p62 protein expression was unchanged ( D ) (Unpaired t-test: p=0,9937). F, G, Western blot representative bands of LC3-I, LC3-II, p62 ( F ) and pAMPK ( G ) in p40 mice and their quantification ( H, I, J ) show no differences between the two genotypes (Unpaired t-test: LC3-II p=0.8446; p62 p=0,7426; pAMPK p=0.6054). Number of mice at p14: LC3-II; Tsc1 Ctrl n=3, Tg( Nkx2.1-Cre ); Tsc1 flox/flox n= 3; p62 and pAMPK Tsc1 Ctrl n = 7, Tg( Nkx2.1-Cre ); Tsc1 flox/flox n= 5. Number of mice at p40: LC3-II and p62; Tsc1 Ctrl n = 4, Tg( Nkx2.1-Cre ); Tsc1 flox/flox n= 4; pAMPK; Tsc1 Ctrl n = 3, Tg( Nkx2.1-Cre ); Tsc1 flox/flox n= 4. Data represent mean ± SEM.

Article Snippet: The following primary antibodies were used: anti-LC3B (1:1000, Novus), anti-p62 (1:500, Proteintech), anti-pAMPK (1:800, Cell Signaling), and anti-GAPDH (1:5000, ThermoFisher).

Techniques: Western Blot, Expressing

Journal: BMC cancer

Article Title: Anti-proliferative activity of RIHMS-Qi-23 against MCF-7 breast cancer cell line is through inhibition of cell proliferation and senescence but not inhibition of targeted kinases.

doi: 10.1186/s12885-023-11547-1

Figure Lengend Snippet: Fig. 4 RIMHS-Qi-23 acts through affecting autophagy and necrosis pathways: a-d Immunoblotting of MCF-7 cell lines homogenates following treatment with serial concentration, incubated with cyclophlin A, p62, LC3 and β-actin as loading control e-g Quantification of immunoblots band values relative to β-actin were normalized to non-treated cells and represent mean ± SEM. Capital letters represent p values from One-Way ANOVA followed by a post-hoc test (similar letters = a statistically non-significant difference, while different letters = a statistically significant difference). Bold values denote significant p values (≤ 0.05)

Article Snippet: Primary antibodies against LC3B (GeneTex catalog no. GTX127375, dilution 1:1000), p62 (SantaCruz Biotechnology catalog no. sc-48389, dilution 1:500), cyclophilin A (Cell signaling catalog no. 2175, dilution 1:1000) and β-actin (Abcam ab227387, dilution 1:5000) were incubated overnight at 4 °C.

Techniques: Western Blot, Concentration Assay, Incubation, Control