Journal:

Article Title: Loss of Oncogenic H-ras-Induced Cell Cycle Arrest and p38 Mitogen-Activated Protein Kinase Activation by Disruption of Gadd45a

doi: 10.1128/MCB.23.11.3859-3871.2003

Figure Lengend Snippet: Disruption of Gadd45a abolishes p38 activation after H-ras overexpression. (A) wt and Gadd45a−/− MEF were infected with H-ras-expressing retrovirus, and activities of ERK, JNK, and p38 were analyzed, with MBP, GST-Jun, and GST-ATF2 as substrates, respectively. puro, puromycin. (B) At the same time that the protein extracts were obtained, the levels of p16/Ink4a, p53, and p21/Waf1 were determined. (C) MEF were infected with either puromycin- or H-ras-expressing retroviruses, and 5 days after selection with puromycin, mRNA was purified and the levels of p53-inducible genes (those encoding p21/Waf1, XPC, and ATF2) were analyzed by using a quantitative filter hybridization procedure (29). The relative induction ratio was obtained after dividing the relative level of mRNA after infection with H-ras retrovirus by the level of mRNA after infection with puromycin vector alone. (D) The levels of Gadd45a and p19/ARF mRNA in wt and Gadd45a−/− MEF on day 5 after retroviral infection were measured by Northern blotting. GAPD was included as a loading control.

Article Snippet: Analysis of protein levels was carried out by using polyclonal antibodies (Abs) against p16/ink4a (M-156; Santa Cruz) and p53 (AB7; Oncogene), monoclonal Abs against p21/Waf1 (AB4; Oncogene) and actin (AB1; Oncogene), anti-Flag monoclonal Ab M2 (Sigma) or anti-Flag monoclonal Ab M2 conjugated with horseradish peroxidase (Sigma), anti-HA polyclonal Ab (Covance Research Products), anti-Myc-tag monoclonal Ab 9E10 (Santa Cruz), anti-Gadd45a polyclonal Ab H-165 (Santa Cruz), anti-p38 polyclonal Ab C-20 (Santa Cruz), anti-JNK polyclonal Ab C-17 (Santa Cruz), and anti-ERK2 polyclonal Ab C-14 (Santa Cruz).

Techniques: Activation Assay, Over Expression, Infection, Expressing, Selection, Purification, Hybridization, Plasmid Preparation, Northern Blot

Journal:

Article Title: Loss of Oncogenic H-ras-Induced Cell Cycle Arrest and p38 Mitogen-Activated Protein Kinase Activation by Disruption of Gadd45a

doi: 10.1128/MCB.23.11.3859-3871.2003

Figure Lengend Snippet: Gadd45a and p38 are required for p53 activation after H-ras overexpression. (A) wt and Gadd45a−/− MEF were incubated in the presence of a MEK1 (50 μM PD98059) or p38 (10 μM SB202190) inhibitor, and protein extracts were obtained on day 5 after selection (see Materials and Methods). The levels of p16/Ink4a, p21/Waf1, and p53 proteins were analyzed. DMSO, dimethyl sulfoxide; puro, puromycin. (B) wt and Gadd45a−/− MEF were cotransfected with p53RE-CAT reporter plasmid and expression vectors containing either puromycin or H-ras. Some cells were additionally transfected with a dominant-negative p38α vector (p38DN). Four days later, cells were treated with either a MEK1 (PD90859) or p38 (SB202190) inhibitor, and CAT assays were carried out 12 h later. (C) wt and Gadd45a−/− MEF were cotransfected with p53RE-CAT reporter plasmid and expression vectors containing either puromycin or MKK6(E). Four days later, CAT activity was analyzed, and representative results are shown. Relative induction, as measured by increased CAT activity, was consistently twofold or greater in wt MEF compared to that in Gadd45a−/− MEF.

Article Snippet: Analysis of protein levels was carried out by using polyclonal antibodies (Abs) against p16/ink4a (M-156; Santa Cruz) and p53 (AB7; Oncogene), monoclonal Abs against p21/Waf1 (AB4; Oncogene) and actin (AB1; Oncogene), anti-Flag monoclonal Ab M2 (Sigma) or anti-Flag monoclonal Ab M2 conjugated with horseradish peroxidase (Sigma), anti-HA polyclonal Ab (Covance Research Products), anti-Myc-tag monoclonal Ab 9E10 (Santa Cruz), anti-Gadd45a polyclonal Ab H-165 (Santa Cruz), anti-p38 polyclonal Ab C-20 (Santa Cruz), anti-JNK polyclonal Ab C-17 (Santa Cruz), and anti-ERK2 polyclonal Ab C-14 (Santa Cruz).

Techniques: Activation Assay, Over Expression, Incubation, Selection, Plasmid Preparation, Expressing, Transfection, Dominant Negative Mutation, Activity Assay

Journal: RSC Advances

Article Title: Design, synthesis, anticancer and in silico assessment of 8-caffeinyl-triazolylmethoxy hybrid conjugates †

doi: 10.1039/d2ra07683g

Figure Lengend Snippet: The cytotoxic activity of 22a–22j and MTX against A-375, MCF7 and MDA-MB-468 and HEK-293 cell lines

Article Snippet: The in vitro evaluations of synthesized compounds on cancer cell lines, including two breast cancer cell lines MDA-MB-468 (ATCC HTB-22), MCF-7 (ATCC HTB-22), melanoma cell line A-375 (ATCC CRL-1619) and normal cell line HEK-293 (ATCC CRL-11268) have determined that 22c (IC 50 < 12.5 μM) demonstrated potent activity against A375 and its toxicity is even stronger than methotrexate (MTX) as a standard drug.

Techniques: Activity Assay

Journal: RSC Advances

Article Title: Design, synthesis, anticancer and in silico assessment of 8-caffeinyl-triazolylmethoxy hybrid conjugates †

doi: 10.1039/d2ra07683g

Figure Lengend Snippet: (A) The viability assessment for 22a–22j and MTX against A375. (B) The viability assessment for 22a–22j and MTX against MCF7. (C) The viability assessment for 22a–22j and MTX against MDA-MB-468. (D) The viability assessment for 22a–22j and MTX against HEK-293.

Article Snippet: The in vitro evaluations of synthesized compounds on cancer cell lines, including two breast cancer cell lines MDA-MB-468 (ATCC HTB-22), MCF-7 (ATCC HTB-22), melanoma cell line A-375 (ATCC CRL-1619) and normal cell line HEK-293 (ATCC CRL-11268) have determined that 22c (IC 50 < 12.5 μM) demonstrated potent activity against A375 and its toxicity is even stronger than methotrexate (MTX) as a standard drug.

Techniques:

Journal: Molecules

Article Title: Do Ganoderma Species Represent Novel Sources of Phenolic Based Antimicrobial Agents?

doi: 10.3390/molecules28073264

Figure Lengend Snippet: Antibacterial activity of H 2 O Ganoderma extracts using three different antibacterial assays.

Article Snippet: G. applanatum showed the highest AB activity against multi-resistant E. coli ATCC 11229 and B. cereus HP with MIC of 12.5 mg/mL.

Techniques: Activity Assay, Diffusion-based Assay

Journal: Molecules

Article Title: Do Ganoderma Species Represent Novel Sources of Phenolic Based Antimicrobial Agents?

doi: 10.3390/molecules28073264

Figure Lengend Snippet: Antibacterial activity of CHCl 3 extracts of G. applanatum and G. resinaceum using microdilution assay.

Article Snippet: G. applanatum showed the highest AB activity against multi-resistant E. coli ATCC 11229 and B. cereus HP with MIC of 12.5 mg/mL.

Techniques: Activity Assay

Journal: Molecules

Article Title: Do Ganoderma Species Represent Novel Sources of Phenolic Based Antimicrobial Agents?

doi: 10.3390/molecules28073264

Figure Lengend Snippet: PCA of quantified phenolic compounds and AB activity of CHCl 3 and H 2 O extracts of G. applanatum and G. resinaceum . Abbreviations: GA— G. applanatum ; GR— G. resinaceum : MBC—minimal bactericidal concentration (mg/mL); MIC—minimal inhibitory concentration (mg/mL); BC— B. cereus ; EC— E. coli ; KP— K. pneumoniae ; PA— P. aeruginosa ; SA— S. aureus .

Article Snippet: G. applanatum showed the highest AB activity against multi-resistant E. coli ATCC 11229 and B. cereus HP with MIC of 12.5 mg/mL.

Techniques: Activity Assay, Concentration Assay

Journal: Current Issues in Molecular Biology

Article Title: Antimicrobial Properties of a Peptide Derived from the Male Fertility Factor kl2 Protein of Drosophila melanogaster

doi: 10.3390/cimb44030076

Figure Lengend Snippet: MFF-p1 is bactericidal in vitro. ( A – C ): E. coli (HB101 strain), B. subtilis (ATCC 9372 strain), and S. aureus (8325-4 strain) were incubated with chemerin-p4, MFF-p1 and Z-p1 (100 μM) peptides for 2 h. Cell viability was analyzed by MDA assay. N = 3, mean ± SEM, ***, and p < 0.001; **, p < 0.01; and *, p < 0.05 by Kruskal–Wallis one-way ANOVA with post hoc Dunn’s test. ( D – F ): Bacteria were incubated with chemerin-p4 or MFF-p1 peptides at the indicated concentrations for 2 h. Data show the percentage of killing (0%—control group). Mean ± SEM of three independent measurements is shown. ****, p < 0.0001; *, p < 0.05 by one-way ANOVA with post hoc Sidak test.

Article Snippet: The selected peptides (100 μM) were tested for antimicrobial activity against E. coli HB101, S. aureus 8325-4 and B. subtilis ATCC 9387 strains using an MDA assay.

Techniques: In Vitro, Incubation, Multiple Displacement Amplification, Bacteria, Control

Journal: Current Issues in Molecular Biology

Article Title: Antimicrobial Properties of a Peptide Derived from the Male Fertility Factor kl2 Protein of Drosophila melanogaster

doi: 10.3390/cimb44030076

Figure Lengend Snippet: MFF-p1 disrupts the integrity of the superficial layers of bacteria. E. coli (HB101 strain) , B. subtilis (ATCC 9372 strain), and S. aureus (8325-4 strain) were incubated with chemerin-p4 (ch-p4) and MFF-p1 (100 μM) peptides for 2 h. Bacteria morphology was assessed by transmission electron microscopy. The images in each panel are from one experiment and are representative of at least three independent experiments.

Article Snippet: The selected peptides (100 μM) were tested for antimicrobial activity against E. coli HB101, S. aureus 8325-4 and B. subtilis ATCC 9387 strains using an MDA assay.

Techniques: Bacteria, Incubation, Transmission Assay, Electron Microscopy

Journal: Current Issues in Molecular Biology

Article Title: Antimicrobial Properties of a Peptide Derived from the Male Fertility Factor kl2 Protein of Drosophila melanogaster

doi: 10.3390/cimb44030076

Figure Lengend Snippet: Gene expression ( A ) and antibacterial protection of MFF kl2 ( B ) in response to microbial challenge. ( A ) Bacteria upregulate the mRNA expression of kl-2 . Canton S flies were infected by injection with E. coli ( E. coli group), injected with PBS (PBS group) or untreated (CTR group) and collected 3 h, 6 h and 9 h after infection, and total RNA was subjected to RT-qPCR. Relative expression of kl-2 is shown as mean ± SD (N = 3 independent experiments, with 15 flies in each). Statistically significant differences between groups are indicated as ** p < 0.01 by two-tailed Student’s t -test. ( B ) Male fertility factor kl2 is bactericidal in vivo. The experimental flies ( act > kl-2 RNAi ) and controls ( act > Valium , w 118 > kl-2 RNAi , act > w 1118 ) were infected with E. coli . Three hours after infection, flies were collected and bacterial load was determined by colony-forming assay. The results are shown as means ± SEM. Statistically significant differences between groups are indicated as ***, p < 0.001; **, p < 0.01; and *, p < 0.05 by Kruskal–Wallis one-way ANOVA with post hoc Dunn’s test.

Article Snippet: The selected peptides (100 μM) were tested for antimicrobial activity against E. coli HB101, S. aureus 8325-4 and B. subtilis ATCC 9387 strains using an MDA assay.

Techniques: Gene Expression, Bacteria, Expressing, Infection, Injection, Quantitative RT-PCR, Two Tailed Test, In Vivo