Journal: International Journal of Molecular Sciences

Article Title: Adjustments of the Phytochemical Profile of Broccoli to Low and High Growing Temperatures: Implications for the Bioactivity of Its Extracts

doi: 10.3390/ijms25073677

Figure Lengend Snippet: The impact of different concentrations of extracts from broccoli cultivated under room (RT), high (HT), and low (LT) temperature on the levels of ROS within ( a ) mouse embryonal fibroblasts (MEF), ( b ) normal human keratinocytes (HaCaT), ( c ) hepatocellular carcinoma (HepG2), ( d ) colorectal carcinoma (HCT116), and ( e ) lung carcinoma (H460) cell cultures. Values represent mean ± standard deviation of three biological replicates. Different letters indicate a significant difference between the RT, HT, and LT broccoli microgreens, separately for each extract concentration (ANOVA, Duncan test, p ≤ 0.05). An asterisk (*) indicates a significant difference between each group of cells treated with broccoli microgreen extracts and control cells (Student’s t -test, p ≤ 0.05). A double asterisk (**) indicates a significant difference between cells treated with H 2 O 2 only and cells treated simultaneously with H 2 O 2 and broccoli extract at a 0.05 mg/mL concentration (Student’s t -test, p ≤ 0.05). AU = arbitrary units; Con = control cells; DCF = dichlorodihydrofluorescein; FI = fluorescence intensity.

Article Snippet: Intracellular ROS levels were determined on mouse embryonal fibroblasts (MEF, established in the laboratory of Dr. Pinteric from mice RRID:IMSR_JAX:002448, Jackson Lab, Bar Harbor, ME, USA), normal human keratinocytes (HaCaT, RRID: CVCL-0038, a gift from Dr. Marijeta Kralj), hepatocellular carcinoma (HepG2, ATCC, HB-8065, Manassas, VA USA), colorectal carcinoma (HCT116, ATCC, CCL-247, Manassas, VA, USA), and lung carcinoma (H460, ATCC, HTB-177, Manassas, VA, USA) cell lines using 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA, cat. no. D6883, Sigma Aldrich, Saint Louis, MO, USA).

Techniques: Standard Deviation, Concentration Assay, Control, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Adjustments of the Phytochemical Profile of Broccoli to Low and High Growing Temperatures: Implications for the Bioactivity of Its Extracts

doi: 10.3390/ijms25073677

Figure Lengend Snippet: The principal component analysis showing ( a ) the relation between broccoli microgreens grown under room (RT), high (HT), and low (LT) temperatures based on the analyzed variables, whose grouping is shown in the ( b ) part of the figure. ABA = abscisic acid; Car = carotenoids; cc = conjugated compound; Chl = chlorophyll; fc = free compound; FerA= ferulic acid; GLS = total intact glucosinolates; H2O2 = hydrogen peroxide; H460 = lung carcinoma; HaCaT = normal human keratinocytes; HCT116 = colorectal carcinoma; HepG2 = hepatocellular carcinoma; IAA = indole-3-acetic acid; K = kaempferol; L-asc = L -ascorbic acid, MEF = mouse embryonal fibroblasts; Porf = porphyrins; PROT = total proteins; Q = quercetin; SinA = sinapic acid; SS = total soluble sugars; TA = total monomeric anthocyanins; TF = total flavonoids; TFlo = total flavonols; THCA = total hydroxycinnamic acids; TP = total phenolics; TPA = total phenolic acids; TPAN = total proanthocyanidins; TT = total tannins.

Article Snippet: Intracellular ROS levels were determined on mouse embryonal fibroblasts (MEF, established in the laboratory of Dr. Pinteric from mice RRID:IMSR_JAX:002448, Jackson Lab, Bar Harbor, ME, USA), normal human keratinocytes (HaCaT, RRID: CVCL-0038, a gift from Dr. Marijeta Kralj), hepatocellular carcinoma (HepG2, ATCC, HB-8065, Manassas, VA USA), colorectal carcinoma (HCT116, ATCC, CCL-247, Manassas, VA, USA), and lung carcinoma (H460, ATCC, HTB-177, Manassas, VA, USA) cell lines using 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA, cat. no. D6883, Sigma Aldrich, Saint Louis, MO, USA).

Techniques:

Journal: PLOS Pathogens

Article Title: Constitutive secretion of pro-IL-18 allows keratinocytes to initiate inflammation during bacterial infection

doi: 10.1371/journal.ppat.1011321

Figure Lengend Snippet: HaCaT, Detroit 562, HEp-2, A-431, primary keratinocytes, or HUVEC cells, were infected with 7.5 x 10 6 colony-forming units (CFU) of GAS for 6 h. (A) Relative abundance of select cytokines was examined by membrane-based antibody array. (B) Cytokine profiles of each cell were examined by multivariate (principal component analysis), of the total variance, PC1 explains 48.67% and PC2 20.87%, from the raw cytokine quantities tabulated in (A). Arrows indicate change in cells from uninfected to 6 h infection. (C) Graphical representation of the congruent cytokine profiles between cell types.

Article Snippet: A-431 lung epidermal cells (ATCC), Detroit 562 pharyngeal cells (ATCC), HaCat cells (unavailable from a repository, provided by C. Quave, Emory), and HEp-2 laryngeal cells (ATCC) were cultured in Eagle’s Minimum Essential Medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (hiFBS, Atlanta Biologicals).

Techniques: Infection, Membrane, Ab Array

Journal: Oncotarget

Article Title: Interaction of microtubules with the actin cytoskeleton via cross-talk of EB1-containing +TIPs and γ-actin in epithelial cells

doi: 10.18632/oncotarget.12236

Figure Lengend Snippet: HaCaT A. - D. or MCF-7 (E) cells were plated for either 6 (A, B, C) or 16 hours (D, E) and stained for β-actin, γ-actin and α-tubulin. Images represent single X/Y sections (A, C, D) and Z section (D, bottom image). Panel B and E represent galleries of optical sections taken with 0.5 μm (B) or 0.3 μm E. step from the ventral (close to the substrate, first image) to the dorsal (last image) side of the HaCaT (B) cell shown in Fig or MCF7 cell (E). Microtubules are distributed in close proximity to the γ-actin network, but not codistributed with the β-actin bundles. Bars, 5 μm (C) and 10 μm.

Article Snippet: Human breast adenocarcinoma MCF7 cell line (ATCC ® HTB22TM) and human immortalized keratinocytes HaCaT (CLS # 300493) were used. pLKO.1 lentiviral DNA constructs together with the pΔR8.2 (#12263, Addgene) and pVSV-G (#8454, Addgene) packaging plasmids were transfected into 293FT packaging cells (R70007, ThermoFisher) using TurboFect Transfection Reagent (R0531, Thermo Scientific).

Techniques: Staining