Journal: Oxidative Medicine and Cellular Longevity

Article Title: Human Tissue Kallikrein 1 Is Downregulated in Elderly Human Prostates and Possesses Potential In Vitro Antioxidative and Antifibrotic Effects in Rodent Prostates

doi: 10.1155/2021/8877540

Figure Lengend Snippet: KLK1 could upregulate NO/cGMP signal and inhibit fibrosis in the prostate of aged rats. (a, b, e, and h) NO content, cGMP content, ADMA content, and cAMP content normalized to total protein concentration of rat prostate samples. (c, f, and i) Representative Western blot results of eNOS, DDAH2, COX-2, PTGIS, TGF- β 1, RhoA, and ROCK1 in prostates of all three groups. (d, g, and j) The expression levels of above-mentioned proteins with β -actin as the loading control in all three groups. For each group, values are presented as the mean ± SD of 10 rats per group. ∗ P < 0.05, ∗∗ P < 0.01 (aTGR vs. aWTR). ## P < 0.01 (aWTR vs. yWTR).

Article Snippet: In Western blot experiments, the primary antibodies against rat KLK1 (Catalog No. ab131029) and human KLK1 (Catalog No. ab28289) were purchased from Abcam (USA); the primary antibodies against NADPH oxidase 2 (NOX2) (Catalog No. 19013-1-AP), NOX4 (Catalog No. 14347-1-AP), p47 phox (Catalog No. 28187-1-AP), p67 phox (Catalog No. 15551-1-AP), Collagen I (Catalog No. 14695-1-AP), Collagen III (Catalog No. 22734-1-AP), α -SMA (Catalog No. 14395-1-AP), DDAH2 (Catalog No. 14966-1-AP), TGF- β 1 (Catalog No. 21898-1-AP), RhoA (Catalog No. 10749-1-AP), ROCK1 (Catalog No. 21850-1-AP), COX-2 (Catalog No. 12375-1-AP), and β -actin (Catalog No. 20536-1-AP) were purchased from Proteintech (China); the primary antibodies against PTGIS (Catalog No. DF4745), inducible NO synthase (iNOS) (Catalog No. AF0199), eNOS (Catalog No. AF0096), β -Tubulin (Catalog No. T0023), and the secondary antibodies were from Affinity (USA).

Techniques: Protein Concentration, Western Blot, Expressing

Journal: Redox Biology

Article Title: A small molecule NRF2 activator BC-1901S ameliorates inflammation through DCAF1/NRF2 axis

doi: 10.1016/j.redox.2020.101485

Figure Lengend Snippet: DCAF1 directs NRF2 for ubiquitination and proteasomal degradation and is the potential target of BC-1901S. A) Immunoblots for NRF2 and β-Actin in Beas-2B cells after knockdown of top-hit E3 ligases JOSD1, BTBD8, or DCAF1 using dsiRNA. B) Immunoblots for DCAF1, NRF2, and β-Actin after DCAF1 knockdown. C) Association between DCAF1 and NRF2 by Co-IP. D) NRF2 Nano-Luc intensity (normalized) after DCAF1 knockdown using two independent dsiRNAs. Data and mean ± SD of 3 independent experiments. E) Immunoblots of DCAF1 and NRF2 expression in after DCAF1 overexpression. F) NRF2 Nano-Luc intensity (normalized) after DCAF1 overexpression in Beas-2B cells. Data and mean ± SD of 3 independent experiments. G) Immunoblots of Beas-2B cells treated with scrambled or DCAF1 dsiRNA followed by IP for NRF2 and IB for K48-Ubiquitin. H) NRF2 Nano-Luc intensity (normalized) in WT, DCAF1-KD, or KEAP1 KO HAP1 NRF2 Nano-Luc cells followed by treatment with MG132 for various time-points. Data and mean ± SD of 3 independent experiments. I) NRF2 binding assay where Beas-2B cells were treated with increasing concentrations of BC-1901S, followed by IP for NRF2 and IB for DCAF1. J) Cell based thermal shift assay (CETSA) of DCAF1 and KEAP1 proteins in Beas-2B cells treated with BC-1901S or control. Blots are representative of 3 independent experiments. K, L) Quantification of DCAF1 and KEAP1 abundance from CETSA experiments. P < 0.05, *; P < 0.0001, ****; by one-way ANOVA with adjusted P-value for multiple comparisons (Dunnett's) compared to control, except indicated specifically.

Article Snippet: Anti-DCAF1 antibody (14966), Anti-HO-1 antibody (mouse specific) (86806), Anti-NRF2 antibody (12721) and K48-linkage Specific Polyubiquitin antibody (8081S) were from Cell signaling technology.

Techniques: Western Blot, Co-Immunoprecipitation Assay, Expressing, Over Expression, Binding Assay, Thermal Shift Assay

Journal: PLoS ONE

Article Title: A New Double Digestion Ligation Mediated Suppression PCR Method for Simultaneous Bacteria DNA-Typing and Confirmation of Species: An Acinetobacter sp. Model

doi: 10.1371/journal.pone.0115181

Figure Lengend Snippet: Reference Acinetobacter strains use in this study.

Article Snippet: Acinetobacter tjernbergiae , DSMZ 14971 , -.

Techniques:

Journal: PLoS ONE

Article Title: A New Double Digestion Ligation Mediated Suppression PCR Method for Simultaneous Bacteria DNA-Typing and Confirmation of Species: An Acinetobacter sp. Model

doi: 10.1371/journal.pone.0115181

Figure Lengend Snippet: Elements of ddLMS PCR typing systems for Acinetobacter sp.

Article Snippet: Acinetobacter tjernbergiae , DSMZ 14971 , -.

Techniques: Polymerase Chain Reaction, Multiplex Assay

Journal: PLoS ONE

Article Title: A New Double Digestion Ligation Mediated Suppression PCR Method for Simultaneous Bacteria DNA-Typing and Confirmation of Species: An Acinetobacter sp. Model

doi: 10.1371/journal.pone.0115181

Figure Lengend Snippet: Electropherogram of PCR products: (A) model system verification, (B) test to verify the specificity of the ddLMS PCR. t- PCR target sequence amplification. pt- PCR – primer-testing PCR; s- PCR specific amplification, ns- PCR nonspecific amplification, cr- PCR cros-check amplification; Three different genomic DNA were used for the ddLSM PCR typing: S , the genomic DNA of A. baumannii ; S+ NS , the genomic DNA of A. baumannii mixed with genomic DNA of other bacteria; and NS , the genomic DNA of bacteria which do not belong to the Acinetobacter genus. M1 the molecular DNA size marker (100–1000 bp); K - , the negative control of the ddLMS PCR (without DNA).

Article Snippet: Acinetobacter tjernbergiae , DSMZ 14971 , -.

Techniques: Sequencing, Amplification, Marker, Negative Control

Journal: PLoS ONE

Article Title: A New Double Digestion Ligation Mediated Suppression PCR Method for Simultaneous Bacteria DNA-Typing and Confirmation of Species: An Acinetobacter sp. Model

doi: 10.1371/journal.pone.0115181

Figure Lengend Snippet: (A) the 3′ recA -ddLMS PCR Mae II/ Rsa I, (B) PCR-RFLP/ Tsp I typing of Acinetobacter calcoaceticus-baumannii complex. Ab1-Ab6 – A. baumannii strains; Ac1-Ac2 – A. calcoaceticus strains; Ap – A. pittii ; An – A. nosocomialis ; 1/3 – Acinetobacter gs. “Between 1 and 3”; C-to-13TU - Acinetobacter gs. “Close-to 13 TU” . M2- the molecular DNA size marker (100-3000 bp); M3 - the molecular DNA size marker (pUC19/ Msp I); K - , the negative control (without DNA).

Article Snippet: Acinetobacter tjernbergiae , DSMZ 14971 , -.

Techniques: Marker, Negative Control

Journal: PLoS ONE

Article Title: A New Double Digestion Ligation Mediated Suppression PCR Method for Simultaneous Bacteria DNA-Typing and Confirmation of Species: An Acinetobacter sp. Model

doi: 10.1371/journal.pone.0115181

Figure Lengend Snippet: (A) 5′ rrn -ddLMS PCR Hind III/ Apa I method and dendrogram generated by Dice Coefficient (DC) and the UPGMA clustering method, (B) PCR MP/ Eco RI typing PCR and dendrogram generated by Dice Coefficient (DC) and the UPGMA clustering method. Ab1–Ab10 – epidemiologically related and unrelated representative Acinetobacter baumannii strains. A–F – genotypes, indicates different fingerprint patterns for unrelated strains. M1- the molecular DNA size marker (100–1000 bp); M2 - the molecular DNA size marker (100–3000); K - , the negative control (without DNA).

Article Snippet: Acinetobacter tjernbergiae , DSMZ 14971 , -.

Techniques: Generated, Marker, Negative Control