Journal: Frontiers in Cell and Developmental Biology

Article Title: Förster Resonance Energy Transfer-Based Single-Cell Imaging Reveals Piezo1-Induced Ca 2+ Flux Mediates Membrane Ruffling and Cell Survival

doi: 10.3389/fcell.2022.865056

Figure Lengend Snippet: Piezo1-mediated extracellular Ca 2+ influx induced by Yoda1 in various cell lines. (A) Viability of MCF-7 cells exposed to control [0.2% (v/v) DMSO] and Yoda1 (0.1–25 μM) for 24 h, measured using the WST-8 assay. The bar graphs describe mean values of cell viability with error bars indicating the standard error of the mean (S.E.M) ( n = 4, ** p < 0.01, student t -test). (B,D) Piezo1 expression in MCF-7 cells with or without control/Piezo1 shRNA as analysed by (B) RT-PCR and (D) western blotting. GAPDH was used as an internal control. (C,E) The bar graphs describe the cumulative densitometric analysis of (C) RT-PCR and (E) western blot bands normalized to the GAPDH. Data are presented as the means ± S.E.M ( n = 4, ** p < 0.01 and *** p < 0.001, student t -test). (F,H) Time-lapse FRET images of the Cytosolic-D3cpv in (F) MCF-7 cells, (H) HEK293A and HeLa cells introduced with or without exogenous Piezo1. MCF-7 cells were exposed to control [0.2% (v/v) DMSO] and Yoda1 (1 μM) dissolved in CO 2 -independent culture medium (Yoda1 group), Ca 2+ -free medium (Yoda1 + w/o Ca 2+ group), and Ca 2+ medium (Yoda1 + w/Ca 2+ group), respectively. (F) Yoda1 + Piezo1 KD group and (H) HEK293A and HeLa cells were incubated in CO 2 -independent culture medium. The color scale bars represent the range of the FRET/CFP emission ratio determined using the biosensor. Hot and cold colors indicate high and low Ca 2+ concentration, respectively. (G,I,J) The time courses represent the average of the normalized FRET/CFP emission ratio changes of cytosolic-D3cpv in (G) MCF-7, (I) HEK293A, and (J) HeLa cells. The lines are mean values of normalized emission ratios, with diluted colors indicating the S.E.M ( n = 7).

Article Snippet: The MCF-7, SiHa, and BeWo cell lines were purchased from the Korean Cell Line Bank (KCLB; Seoul, Republic of Korea), and HEK293A and HeLa cell lines were provided by Dr. Jihye Seong (Korea Institute of Science and Technology, Seoul, South Korea).

Techniques: Expressing, shRNA, Reverse Transcription Polymerase Chain Reaction, Western Blot, Incubation, Concentration Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Förster Resonance Energy Transfer-Based Single-Cell Imaging Reveals Piezo1-Induced Ca 2+ Flux Mediates Membrane Ruffling and Cell Survival

doi: 10.3389/fcell.2022.865056

Figure Lengend Snippet: Piezo1-dependent ER-stored Ca 2+ release in response to Yoda1. (A,D) Time-lapse FRET images of the ER Ca 2+ sensor in (A) MCF-7 and (D) HeLa cells. The cells were exposed to control [0.2% (v/v) DMSO] and Yoda1 (1 μM) dissolved in CO 2 -independent culture medium (Yoda1 group), Ca 2+ -free medium (Yoda1 + w/o Ca 2+ group), and Ca 2+ medium (Yoda1 + w/Ca 2+ group), respectively. The color scale bars represent the range of the FRET/CFP emission ratio determined using the biosensor. Hot and cold colors indicate high and low Ca 2+ concentration, respectively. (D) The red fluorescence protein images confirmed the expression of Piezo1-tdTomato. (B,E) The time courses represent the average of the normalized FRET/CFP emission ratio changes of the ER Ca 2+ sensor. The lines are mean values of the normalized emission ratios, with diluted colors indicating the S.E.M ( n = 7). (C,F) The bar graph describes the mean values of the normalized FRET/CFP emission ratios of the ER Ca 2+ sensor at the described time with error bars indicating the S.E.M ( n = 7, * p < 0.05 and ** p < 0.01, Student’s t -test).

Article Snippet: The MCF-7, SiHa, and BeWo cell lines were purchased from the Korean Cell Line Bank (KCLB; Seoul, Republic of Korea), and HEK293A and HeLa cell lines were provided by Dr. Jihye Seong (Korea Institute of Science and Technology, Seoul, South Korea).

Techniques: Concentration Assay, Fluorescence, Expressing

Journal: Nanomaterials

Article Title: Anticancer Potential of L-Histidine-Capped Silver Nanoparticles against Human Cervical Cancer Cells (SiHA)

doi: 10.3390/nano11113154

Figure Lengend Snippet: ( I ) Antiproliferative effect of L-HAgNPs against SiHa cells; ( II ) phase-contrast microscopic images to assess structural changes in cells treated with L-HAgNPs; ( III ) fluorescent images illustrating intracellular ROS level; ( IV ) bar graph depicting quantitative measurement of green fluorescent intensity proportional to ROS level. * p < 0.05 denotes statistical significance between control vs. treated groups.

Article Snippet: SiHa cells were purchased from National Centre for Cell Science (NCCS), Pune, India.

Techniques: Control