Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Role for polo-like kinase 4 in mediation of cytokinesis

doi: 10.1073/pnas.1818820116

Figure Lengend Snippet: Western immunoblot of 19 different ovarian cell lines and one colorectal (NCI-H747) cancer cell line demonstrates full-length PLK4 ( Upper Left and Upper Right ) and phospho-PLK4 ( Lower Left and Lower Right ). ( Upper ) Western blots demonstrate full-length (97-kDa) PLK4 with a commercially available anti-PLK4 antibody (P005; Cell Signaling 3258, recognizing an epitope centered on the cysteine458 amino acid). ( Lower ) Western blots were performed by using the same filter after washing with buffer and reprobing with an anti–phospho-serine305-PLK4 antibody (Ab#14299). Please note that the phospho-PLK4 in the lower filter is of substantially smaller size, ∼60 kDa. The difference is appreciated as the full-length signals were not completely removed by stripping in the lower filters. Cultured cells were not synchronized with regard to cell cycle. We confirmed that the 60-kDa band corresponded to pPLK4 by removing this band from the gel and performing amino acid sequence analyses. We also confirmed that a second anti–phospho-PLK4 antibody (Ab#3) from our laboratory recognized phospho-PLK4 by using immunoprecipitation assays followed by PAGE with silver staining to identify the band recognized on Western blot and, finally, MS to assess the amino-acid sequence of protein band (amino-acid sequence provided in ). SI Appendix , Fig. S1 shows results in 23 additional colorectal and 9 breast cancer cell lines. ( Lower ) Schematic illustration of PLK4 amino-acid sequence shows the relative locations of epitopes recognized by the two antibodies used for Western immunoblot analyses ( Upper ), one from the laboratory of G.P.N. recognizing phospho-serine305-PLK4, produced in collaboration with Cell Signaling Technology, and the second commercially available recognizing cysteine458-PLK4.

Article Snippet: We used breast cancer (HCC1569, HCC1954, BT549, MDA-MB-415, HCC202, MCF7, T47D, CAL51), ovarian cancer (KOC-7C, SKOV3, HEYC2, COLO704, RMG-1, KK, ES-2, EFO21, MCAS, A2780, OV167, PEA2, TOV-21G, OV90, OV207, Kuramochi, OVISE, OVMANA, PEO6, CaOv-3, OV177, OVCAR3, OVCAR5, OVSAHO, OVTOKO, PEO14), colorectal cancer (HCT-116, HCT-15, DLD-1, COLO 205, COLO320DM, COLO320HSR, NCI-H747, NCI-H716, COLO201, LS1034, SNU-C2B, SNU-C1, LS513, LS411N, ATR-FLOX, LS174t, WiDr, RKO, SK-CO1, HCT-8, LS-180, HT-29, C2Bbe1, SW480), and immortalized breast epithelial (MCF-10–2A, MCF-12A) cell lines from the American Type Culture Collection (ATCC) or the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ) for our experiments grown as described elsewhere ( – ).

Techniques: Western Blot, Stripping Membranes, Cell Culture, Sequencing, Immunoprecipitation, Silver Staining, Produced

Journal: Frontiers in Pharmacology

Article Title: Dehydroepiandrosterone suppresses human colorectal cancer progression through ER stress-mediated autophagy and apoptosis in a p53-independent manner

doi: 10.3389/fphar.2024.1464647

Figure Lengend Snippet: DHEA suppresses the growth and colony formation of human CRC cells regardless of p53 status. (A) IC 50 values of DHEA of five human CRC cell lines, including wild-type p53 (HCT116 p53 +/+ and RKO), mutant p53 (LS1034 and HT29), and null p53 (HCT116 p53 −/− ) cells were determined after 48 and 72 h of DHEA treatment. The IC 50 values were determined by measuring the optical density (OD) at wavelength 510 nm using SRB assay. Statistical analysis was conducted by comparing with HCT116 p53 +/+ ; ns: non-significance with p-value > 0.05, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. (B) The percentage of inhibition of colony formation by DHEA, ranging from 0 to 200 μM, was determined after 12 days of treatment on HCT116 p53 +/+ , HT29, and HCT116 p53 −/− cells. Statistical analysis was conducted to compare the effect of DHEA between treatment groups and the untreated group of HCT116 p53 +/+ , HT29, and HCT116 p53 −/− cells and between 3 cells. Statistical data were obtained from three independent experiments with ns: non-significant with p-value > 0.05, ****p < 0.0001; Error bar is Standard Error of Mean (SEM).

Article Snippet: LS1034 cells were obtained from the Bioresource Collection and Research center (BCRC), Taiwan, and were cultured in Roswell Park Memorial Institute (RPMI 1640) medium.

Techniques: Mutagenesis, Sulforhodamine B Assay, Inhibition

Journal:

Article Title: Biodegradation of Methyl tert -Butyl Ether by a Pure Bacterial Culture

doi: 10.1128/AEM.67.12.5601-5607.2001

Figure Lengend Snippet: Growth of ENV735 on TBA + 0.01% YE (▪), MTBE + 0.01% YE (●), and 0.01% YE (○). Cell growth on MTBE as a sole carbon source was slow and resulted in the formation of dense clumps that were difficult to sample and quantify.

Article Snippet: H. flava ENV735 (ATCC PTA-2158) was isolated by enrichment culturing on MTBE ( 29 ).

Techniques:

Journal:

Article Title: Biodegradation of Methyl tert -Butyl Ether by a Pure Bacterial Culture

doi: 10.1128/AEM.67.12.5601-5607.2001

Figure Lengend Snippet: Degradation of MTBE (A) and TBA (B) by ENV735. Cells were grown on either LB (▾), 0.4% YE (▪), TBA (⧫), CSL (▴), or sucrose (▵) and were assayed for MTBE and TBA biodegradation as described in Materials and Methods. Samples containing poisoned LB-grown cells are also represented (●). These samples are not shown in panel B because no TBA was produced in the samples.

Article Snippet: H. flava ENV735 (ATCC PTA-2158) was isolated by enrichment culturing on MTBE ( 29 ).

Techniques: Produced

Journal:

Article Title: Biodegradation of Methyl tert -Butyl Ether by a Pure Bacterial Culture

doi: 10.1128/AEM.67.12.5601-5607.2001

Figure Lengend Snippet: Biodegradation of MTBE (A) and TBA (B) by strain ENV735 in the presence of known metabolic inhibitors and inactivators. ○, no inhibitor; □, CO; ▿, ATU; ●, ABT; ▪, butadiene monoepoxide; and ▾, acetylene. Values represent means of triplicate samples, and error bars represent 1 standard deviation of the mean.

Article Snippet: H. flava ENV735 (ATCC PTA-2158) was isolated by enrichment culturing on MTBE ( 29 ).

Techniques: Standard Deviation

Journal:

Article Title: Biodegradation of Methyl tert -Butyl Ether by a Pure Bacterial Culture

doi: 10.1128/AEM.67.12.5601-5607.2001

Figure Lengend Snippet: Effect of formaldehyde on MTBE (A) and TBA (B) degradation by ENV735. Symbols in panel A are as follows: ●, MTBE only; ○, TBA produced in MTBE-only samples; ▪, MTBE + 1.3 mM formaldehyde; □, TBA produced in MTBE + 1.3 mM formaldehyde samples; ▴, MTBE + 2.6 mM formaldehyde; and ▵, TBA produced in MTBE + 2.6 mM formaldehyde samples. Symbols in panel B represent mean ± standard deviation (n = 3) of TBA concentration after 20 h of incubation with different concentrations of formaldehyde.

Article Snippet: H. flava ENV735 (ATCC PTA-2158) was isolated by enrichment culturing on MTBE ( 29 ).

Techniques: Produced, Standard Deviation, Concentration Assay, Incubation

Journal:

Article Title: Biodegradation of Methyl tert -Butyl Ether by a Pure Bacterial Culture

doi: 10.1128/AEM.67.12.5601-5607.2001

Figure Lengend Snippet: SDS-PAGE gel of ENV735 proteins after growth on different substrates. Lanes 1 and 2, LB medium; lanes 3 and 6, TBA + 0.01% YE; lanes 4 and 5, MTBE + 0.01% YE; and lane 7, protein size markers. Protein size in kilodaltons is shown to the right of the figure. Arrows A and B identify peptides produced during growth on MTBE and TBA but not during growth on LB.

Article Snippet: H. flava ENV735 (ATCC PTA-2158) was isolated by enrichment culturing on MTBE ( 29 ).

Techniques: SDS Page, Produced

Journal:

Article Title: Biodegradation of Methyl tert -Butyl Ether by a Pure Bacterial Culture

doi: 10.1128/AEM.67.12.5601-5607.2001

Figure Lengend Snippet: Phosphorimage of 14C-labeled proteins from ENV735. TBA-grown cells were incubated with uniformly labeled [14C]MTBE in the presence (lanes 2, 3, 5, and 6) or absence (lanes 1, 4, and 7) of chloramphenicol and were then denatured and separated on a PAGE gel. The gel was dried and placed on a phosphorimaging system to identify 14C-labeled proteins. Protein size (41 kDa) is indicated by arrow.

Article Snippet: H. flava ENV735 (ATCC PTA-2158) was isolated by enrichment culturing on MTBE ( 29 ).

Techniques: Labeling, Incubation