Journal: Journal of Bacteriology

Article Title: The MerR-Like Transcriptional Regulator BrlR Contributes to Pseudomonas aeruginosa Biofilm Tolerance

doi: 10.1128/JB.00765-12

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

Article Snippet: Where necessary, Escherichia coli cultures were grown in LB broth in the absence or presence of 1 mM isopropyl-β- d -thiogalactopyranoside (IPTG). table ft1 table-wrap mode="anchored" t5 caption a7 Strain or plasmid Relevant genotype or description Source or reference Escherichia coli DH5α F − φ80d lacZ ΔM15 Δ( lacZYA - argF ) U169 recA1 endA1 hsdR17 (r K − m K + ) phoA supE44 thi - 1 gyrA96 relA1 tonA Invitrogen Corp. BL21 F − ompT hsdS B (r B − m B − ) gal dcm rne131 (DE3) Invitrogen Corp. P. aeruginosa PAO1 PAO1 Wild type B. H. Holloway PAO1/pJN105 PAO1 bearing empty pJN105 vector; Gm r 55 PAO1/pMJT1 PAO1 bearing empty pMJT1 vector; Carb r This study PAO1/pJN- brlR Arabinose-inducible expression of brlR ; Gm r This study PAO1/pMJT- brlR -V5/6×His Arabinose-inducible expression of V5/6×His-tagged brlR ; Carb r This study Δ brlR mutant PAO1 Δ brlR (PA4878) This study Δ brlR /pJN- brlR mutant Arabinose-inducible expression of brlR ; Gm r This study Δ brlR /pMJT1 mutant Δ brlR harboring empty pMJT1, vector control; Carb r This study Δ brlR /pMJT- brlR -N mutant Arabinose-inducible expression of N-terminal DNA binding domain of BrlR (BrlR-N) in Δ brlR ; Carb r This study CF1-2 Isolate from CF patient 53 CF1-8 Isolate from CF patient 53 CF1-8/pJN- brlR Arabinose-inducible expression of brlR in CF1-8; Gm r This study CF1-8 Δ brlR CF1-8 harboring Δ brlR (PA4878), Gm r This study CF1-13 Isolate from CF patient 53 CF1-13 Δ brlR CF1-13 harboring Δ brlR (PA4878); Gm r This study PAO1 attB :: lacZ pminiCTX- lacZ conjugated into PAO1, vector control; Tet r This study PAO1 attB ::PbrlR- lacZ pminiCTX-PbrlR- lacZ conjugated into PAO1; Tet r This study Plasmids pCR2.1-TOPO TA cloning vector; Km r Ap r Invitrogen Corp. pRK2013 Helper plasmid for triparental mating; mob tra Km r 25 pEX18Gm Gene replacement vector; pUC18 MCS oriT sacB Gm r 26 pJN105 Arabinose-inducible gene expression vector; pBRR-1 MCS araC -P BAD Gm r 50 pMJT1 Arabinose-inducible gene expression vector; pUCP18 MCS araC -P BAD Amp/Carb r 35 pET101D Vector for directional cloning and high level V5/6×His fusion protein expression; Amp r Invitrogen Corp. pminiCTX- lacZ attB site-specific integration vector; Tet r 10 pET- brlR -V5/6×His brlR cloned into pET101D This study pJN- brlR brlR cloned into pJN105 This study pMJT- brlR -V5/6×His brlR -V5/6×His cloned into pMJT1 This study pMJT- brlR -N N-terminal DNA binding domain of BrlR cloned into pMJT1 This study pminiCTX-PbrlR- lacZ brlR promoter reporter construct in mini-CTX- lacZ using PbrlR-lacZF/R primers; Tet r This study Open in a separate window Bacterial strains and plasmids used in this study

Techniques: Plasmid Preparation, Expressing, Mutagenesis, Control, Binding Assay, TA Cloning, Gene Expression, Cloning, Clone Assay, Construct

Journal: Cell reports

Article Title: tRNA methylation resolves codon usage bias at the limit of cell viability

doi: 10.1016/j.celrep.2022.111539

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The gene together with a partial sequence of P BAD was isolated from pACYC- araC -P C -P BAD - EctrmD ( ; ) by excision with BamHI and XhoI (NEB) and gel purified by NucleoSpin Gel and PCR Clean-up (Macherey-Nagel).

Techniques: Recombinant, Clone Assay, Luciferase, Sequencing, Plasmid Preparation, CRISPR, Mutagenesis, Northern Blot, Software, Imaging

Journal: British Journal of Cancer

Article Title: Anti-Cripto Mab inhibit tumour growth and overcome MDR in a human leukaemia MDR cell line by inhibition of Akt and activation of JNK/SAPK and bad death pathways

doi: 10.1038/sj.bjc.6603641

Figure Lengend Snippet: Inhibition of CEM/A7R cells by combined use of anti-Cripto Mab C13 with different concentrations of EPI at 0.11 to 1 μ g ml −1 ( A ), DAU at 0.04–0.33 μ g ml −1 ( B ) or C4 at various concentrations with Arac at 0.0015, 0.003 and 0.005 μ g ml −1 ( C ) after 48 h incubation as measured by percentage of control in [ 3 H]thymidine incorporation. Points are means of triplicate experiments. Error bars represent s.d. in triplicate experiments. The interactions of various concentrations of C13 with EPI, DAU or C4 with AraC were subjected to two-way ANOVA. The P- values were marked as * P <0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Daunorubicin, EPI and cytosine arabinoside (AraC) were obtained from Pharmacia & Upjohn Pty Limited (Perth, Australia).

Techniques: Inhibition, Incubation, Control

Journal: British Journal of Cancer

Article Title: Anti-Cripto Mab inhibit tumour growth and overcome MDR in a human leukaemia MDR cell line by inhibition of Akt and activation of JNK/SAPK and bad death pathways

doi: 10.1038/sj.bjc.6603641

Figure Lengend Snippet: P-glycoprotein function and expression affected by anti-Cripto Mab in CCRF-CEM and CEM-A7R tested by flow cytometry and Western blot. ( A and C ) Flow cytometry analysis of Rh123 accumulation in the absence (open histogram) or presence of 10 μ M Vp (solid histogram). ( B ) Rh123 accumulation in the CEM/A7R cells following 3 h treatment with 25 μ g ml −1 C13 in the absence (open histogram) or presence of 10 μ M Vp (solid histogram) or in the absence (open histogram) or presence of 25 μ g ml −1 C13 (solid histogram) ( D ). P-glycoprotein function is expressed as the ratio of MCF in the presence or absence of Vp or C13 as described in the ‘Materials and Methods’. P-glycoprotein levels were expressed as the ratio (R) of the MCF. ( E ) Western blot analysis of Pgp expression detected by Mab to human Pgp in the CEM/A7R cells following 6 h treatment with 10 μ g ml −1 C4, 0.02 μ g ml −1 AraC and combined use of the two reagents.

Article Snippet: Daunorubicin, EPI and cytosine arabinoside (AraC) were obtained from Pharmacia & Upjohn Pty Limited (Perth, Australia).

Techniques: Expressing, Flow Cytometry, Western Blot

Journal: British Journal of Cancer

Article Title: Anti-Cripto Mab inhibit tumour growth and overcome MDR in a human leukaemia MDR cell line by inhibition of Akt and activation of JNK/SAPK and bad death pathways

doi: 10.1038/sj.bjc.6603641

Figure Lengend Snippet: Western blot showing signalling molecules mediated by C4, AraC and combination of the two reagents in the CEM/A7R cells. ( A ) Activation of JNK/SAPK examined by JNK kinase assay and blotted by anti- phosphor-c-Jun Mab in CEM/A7R cells treated with 10 μ g ml −1 C4 and 0.02 μ g ml −1 AraC or combination for 3 or 6 h. ( B ) Western blot analysis of phospho-Akt at Ser473 and cleavage of caspase 9 at Asp330 compared to the level of expression of Cripto and ( C ) Western blot analysis of phosphor-Bad at Ser136, Ser112, total Bad protein, phosphor-Bcl-2 at Ser70, Bcl-xL in the same cell lysates.

Article Snippet: Daunorubicin, EPI and cytosine arabinoside (AraC) were obtained from Pharmacia & Upjohn Pty Limited (Perth, Australia).

Techniques: Western Blot, Activation Assay, Kinase Assay, Expressing