Journal: British Journal of Pharmacology

Article Title: Inhibition of transmembrane member 16A calcium‐activated chloride channels by natural flavonoids contributes to flavonoid anticancer effects

doi: 10.1111/bph.13841

Figure Lengend Snippet: The effects of flavonoid compounds on recombinant TMEM16A currents in CHO cells. (A) Chemical structures of flavonoids. (B) Chemical structures of tannic acid. (C) The time course for the effects of luteolin (Lute), galangin (Gala), quercetin (Quer) and fisetin (Fise), all at 100 μM and DMSO (0.1%) on TMEM16A currents tested at +100 mV. The protocol was shown at the top of the figure. (D) The representative current traces recorded when the effect of drugs has stabilized. (E) The inhibition by flavonoids (100 μM), DMSO (0.1%), tannic acid, T16Ainh‐A01 and CaCCinh‐A01 (100 μM) of TMEM16A current tested at +100 mV. NS, not significant; *P < 0.05, significant effect of treatments.

Article Snippet: CHO cells, LA795 cells, HEK293 cells and normal lung 2BS cells were cultured in F‐12K (Solarbio, China) (1% nonessential amino acids, 600 μg·mL −1 G418), RPMI 1640 (Solarbio, China) or DMEM medium (Solarbio, China) supplemented with 10% fetal bovine serum (Gibco, USA) and antibiotics (100 IU·mL −1 penicillin G and 100 mg·mL −1 streptomycin; Solarbio, China) in a humidified incubator at 37°C and 5% CO 2 .

Techniques: Recombinant, Inhibition

Journal: British Journal of Pharmacology

Article Title: Inhibition of transmembrane member 16A calcium‐activated chloride channels by natural flavonoids contributes to flavonoid anticancer effects

doi: 10.1111/bph.13841

Figure Lengend Snippet: The effects of luteolin (Lute), galangin (Gala), quercetin (Quer) and fisetin (Fise) on I–V relationship and deactivation kinetics of TMEM16A currents in CHO cells. (A) Representative traces of TMEM16A currents recorded using the voltage protocol indicated at the top of B. Dotted lines indicated the zero current level. The effects of luteolin, galangin, quercetin and fisetin (all at 100 μM) or tannic acid (100 μM) are shown. (B) Normalized current–voltage relationships of TMEM16A currents in the absence or presence of the compounds; n = 5 for each experimental group. (C) The effects of luteolin, galangin, quercetin and fisetin on the deactivation kinetics of TMEM16A currents from +100 to −100 mV. The red line shows the deactivating currents in the presence of the compounds, which were scaled up to match the amplitude of the deactivating currents in the absence of the compounds. (D) Summary of effects of luteolin, galangin, quercetin and fisetin on the time constants of TMEM16A deactivating currents. *P < 0.05, significantly different from control, n = 5 for each experimental group.

Article Snippet: CHO cells, LA795 cells, HEK293 cells and normal lung 2BS cells were cultured in F‐12K (Solarbio, China) (1% nonessential amino acids, 600 μg·mL −1 G418), RPMI 1640 (Solarbio, China) or DMEM medium (Solarbio, China) supplemented with 10% fetal bovine serum (Gibco, USA) and antibiotics (100 IU·mL −1 penicillin G and 100 mg·mL −1 streptomycin; Solarbio, China) in a humidified incubator at 37°C and 5% CO 2 .

Techniques: Control

Journal: British Journal of Pharmacology

Article Title: Inhibition of transmembrane member 16A calcium‐activated chloride channels by natural flavonoids contributes to flavonoid anticancer effects

doi: 10.1111/bph.13841

Figure Lengend Snippet: The concentration–response relationships for luteolin (Lute), galangin (Gala), quercetin (Quer) and fisetin (Fise) on recombinant TMEM16A currents in CHO cells. (A) The time course for the effects of galangin (1–100 μM) on TMEM16A currents tested at +100 mV. (B) The representative current traces recorded when the effect of different concentrations of galangin had stabilized. (C) Concentration–response relationships for luteolin, galangin, quercetin and fisetin on TMEM16A currents recorded at +100 mV. Data were fitted with logistic function. Luteolin: n = 5, 5, 5, 5 and 8; galangin: n = 5, 5, 9, 5 and 7; quercetin: n = 5, 5, 5, 7 and 9; fisetin: n = 5, 5, 5, 6 and 9 (1, 3, 10, 30 and 100 μM).

Article Snippet: CHO cells, LA795 cells, HEK293 cells and normal lung 2BS cells were cultured in F‐12K (Solarbio, China) (1% nonessential amino acids, 600 μg·mL −1 G418), RPMI 1640 (Solarbio, China) or DMEM medium (Solarbio, China) supplemented with 10% fetal bovine serum (Gibco, USA) and antibiotics (100 IU·mL −1 penicillin G and 100 mg·mL −1 streptomycin; Solarbio, China) in a humidified incubator at 37°C and 5% CO 2 .

Techniques: Concentration Assay, Recombinant

Journal: Microorganisms

Article Title: Emergence and Characterization of Three Pseudorabies Variants with Moderate Pathogenicity in Growing Pigs

doi: 10.3390/microorganisms13040851

Figure Lengend Snippet: Isolation, identification, and biological characteristics of the three isolated field PRV strains. ( A ) Vero cells were inoculated with SD1501, SD1701, and SD1801, and the cytopathic effects and fluorescence were observed. ( B ) Identification of SD1501, SD1701, and SD1801 using Western blot analysis. ( C ) Plaque purification of SD1501, SD1701, and SD1801. ( D , E ) Growth curves of SD1501, SD1701, and SD1801 using qPCR ( D ) and TCID50 determination ( E ).

Article Snippet: Vero cells were cultured in Dulbecco’s modified Eagle medium (DMEM) (Solarbio, Shanghai, China), supplemented with 10% fetal bovine serum (FBS, NULEN BIOTECH, Shanghai, China), and maintained at 37 °C in 5% CO 2 .

Techniques: Isolation, Fluorescence, Western Blot, Purification

Journal: Clinical Cancer Research

Article Title: Anti-EGFR Antibody–Drug Conjugate Carrying an Inhibitor Targeting CDK Restricts Triple-Negative Breast Cancer Growth

doi: 10.1158/1078-0432.CCR-23-3110

Figure Lengend Snippet: Stochastic conjugation of cetuximab to CDK inhibitor and ADC internalization in live breast cancer cells. A, Flow cytometric evaluation of surface EGFR expression (TNBC: MDA-MB-468, HCC1143, HCC1806, MDA-MB-231, HCC1937, SUM149, and CAL51; HER2+: SKBR3; ER+: MCF7, T47D; nontumorigenic epithelial cell model: MCF10A; immune cell model: human B lymphocytes RPMI8866, RPMI8226, and monocytic cell line U937; human primary melanocyte: melanocyte). B, EGFR mRNA expression from the Cancer Cell Line Encyclopedia database showed a positive correlation with surface EGFR measured by flow cytometry in A (Spearman’s rank coefficient, r = 0.723). A high level of correlation was found between EGFR and cyclin E ( r = 0.738), but not with cyclin A or CDK2. Nonsignificant P values are marked as NS. C, Top, Schematic diagram of stochastic ADC conjugation by antibody reduction with TCEP and then conjugation to SNS-032 via MC-Val–Ala-PAB. Middle, HIC analysis confirmed an average DAR of 4.4. Bottom, SEC trace indicates negligible ADC aggregation and minimal free linker–payload (less than 0.8%). D, Surface plasmon resonance analysis demonstrated similar binding affinity ( K D ) for cetuximab (0.73 nmol/L) and ADC (1.28 nmol/L). Isotype IgG1 and isotype ADC showed no measurable binding. E, Monitoring internalization of Fabfluor-pH-labeled cetuximab, ADC, or isotype control (10 nmol/L) by Incucyte live-cell imaging. Phase and red fluorescence time-course images were captured for 24 hours. Images of internalized antibody display in cytosolic, low pH lysosomal vesicle-associated red fluorescence in cells. Scale bar, 0.2 mm. F, Cells were seeded in Matrigel for 5 days, allowing the formation of spheroids. Fabfluor-pH-labeled antibodies or ADC (10 nmol/L) were introduced in the Matrigel and showed rapid internalization in EGFR-high MDA-MB-468 and MDA-MB-231, whereas EGFR-low CAL51 displayed little red fluorescence signals. A low level of internalization was observed for isotype or isotype-ADC controls. Scale bar, 0.5 mm. P values determined by two-tailed unpaired t test of three independent experiments compared with isotype control.

Article Snippet: Human primary epidermal melanocytes were cultured in Melanocyte Growth Medium (Cell Applications Inc.).

Techniques: Conjugation Assay, Expressing, Flow Cytometry, SPR Assay, Binding Assay, Labeling, Control, Live Cell Imaging, Fluorescence, Two Tailed Test