Journal: PLoS ONE

Article Title: Nesprins: Tissue-Specific Expression of Epsilon and Other Short Isoforms

doi: 10.1371/journal.pone.0094380

Figure Lengend Snippet: Nesprin DV23 inclusion in cDNA from tissues and cells.

Article Snippet: Ntera-2 (pluripotent neuroectodermal human testicular embryonic teratocarcinoma cell line, gift from Peter Andrews, Sheffield University ), LCL (lymphoblastoid cell line ), HeLa (human epithelial carcinoma cell line ), fibroblasts (human fibroblasts established in culture from skin biopsy ), and U2OS (human osteosarcoma epithelial cell line, obtained from American Tissue Culture Collection (ATCC)), were grown in DMEM with 10% fetal bovine serum and antibiotics and VSMC (vascular smooth muscle cells ) were grown in Medium 199 with 20% fetal bovine serum and antibiotics.

Techniques:

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: IPI measurement of LDs in U2OS cells. (A) Single-color (left) and two-color (right) IP images at 2193 cm −1 and 2855 cm −1 . The two-color IP image was constructed by superimposing IP images simultaneously obtained with IR excitations of 2855 cm −1 (45 kHz) and 2193 cm −1 (50 kHz), which are marked by false green and red colors, respectively. The IPI contrasts were adjusted to reveal the cellular structures. The inset image is the bright-field image of the corresponding region. Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B) IP spectra of LDs before and after background calibration. Three LDs are indicated in . Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. The spectra on the bottom were calibrated by matching the background signal level in the off-resonant spectral region (2740 cm −1 to 2760 cm −1 ). Stepsize, 1 cm −1 . (C) Histogram for IP intensity at 2855 cm −1 of LDs. We obtained IP spectra of 47 LDs within U2OS cells from seven different regions, and the histogram shows the distribution of IP intensities at 2855 cm −1 of the calibrated IP spectra.

Article Snippet: U2OS cell line (human bone osteosarcoma epithelial cells) and Huh-7 cell line (human hepatocellular carcinoma cells) were supplied from Korean Cell Line Bank (Seoul, Korea) and cultured in DMEM (Welgene, LM 001-17) supplemented with penicillin (100 U mL −1 ), streptomycin (100 μg mL −1 ), 10% heat-inactivated fetal bovine serum (Welgene, S1010-01) at 37 °C and pH 7.4 (5% CO 2 ).

Techniques: Construct

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: Two-color IPI measurements of U2OS cells cultured exposed to PA and PA-d 31 . (A) Two-color IP images of fixed U2OS cells cultured in different growth media (250 μM PA, 250 μM PA-d 31 , and standard medium) for 24 hours. The green and red false colors indicate IPI contrasts of 2855 cm −1 and 2193 cm −1 , respectively. Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bar, 20 μm. (B) Representative IP spectra of LDs observed in at two different spectral regions. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Stepsize, 1 cm −1 .

Article Snippet: U2OS cell line (human bone osteosarcoma epithelial cells) and Huh-7 cell line (human hepatocellular carcinoma cells) were supplied from Korean Cell Line Bank (Seoul, Korea) and cultured in DMEM (Welgene, LM 001-17) supplemented with penicillin (100 U mL −1 ), streptomycin (100 μg mL −1 ), 10% heat-inactivated fetal bovine serum (Welgene, S1010-01) at 37 °C and pH 7.4 (5% CO 2 ).

Techniques: Cell Culture

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: Investigation of the LDs with time. (A) Two-color (excitation: 2193 cm −1 and 2855 cm −1 ) IP images of fixed U2OS cells at different time points after PA-d 31 administration. PA-d 31 treatment times were indicated in the upper left corner of each image. The color scheme is the same as in . Cutoff frequency, 70 Hz. Step size, 200 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B) IP spectra of LDs at time points (20 min, 1 h, 6 h, and 24 h) after PA-d 31 administration. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Stepsize, 1 cm −1 . The y -axis scales in the two regions (2030 cm −1 to 2230 cm −1 and 2750 cm −1 to 2950 cm −1 ) are adjusted differently to enhance clarity. (C) Two-color IP images of fixed U2OS cells treated with PA-d 31 for 3 hours. The experimental condition is the same as that of . Scale bars, 20 μm. The figures below depict magnified images of five LDs with 2 μm scale bars. (D) Mole fractions of CD 2 and CH 2 groups from neutral lipids of . Two wavelengths of 2193 cm −1 and 2855 cm −1 were chosen to calculate the mole fraction. (E) Temporal change in mole fraction. The mole fraction of 10 different LDs was averaged at each time point. The error bars represent the upper and lower bounds of the mole fraction, corresponding to the maximum and minimum values, respectively.

Article Snippet: U2OS cell line (human bone osteosarcoma epithelial cells) and Huh-7 cell line (human hepatocellular carcinoma cells) were supplied from Korean Cell Line Bank (Seoul, Korea) and cultured in DMEM (Welgene, LM 001-17) supplemented with penicillin (100 U mL −1 ), streptomycin (100 μg mL −1 ), 10% heat-inactivated fetal bovine serum (Welgene, S1010-01) at 37 °C and pH 7.4 (5% CO 2 ).

Techniques:

Journal: Chemical Science

Article Title: Monitoring the synthesis of neutral lipids in lipid droplets of living human cancer cells using two-color infrared photothermal microscopy †

doi: 10.1039/d3sc04705a

Figure Lengend Snippet: Real-time monitoring of neutral lipid synthesis in living U2OS cells. (A) Time-lapse two-color (excitation: 2193 cm −1 and 2855 cm −1 ) IP images of a live U2OS cell. The two-color IP images were obtained every 10 minutes for 4 hours. The color scheme is the same as in . Cutoff frequency, 70 Hz. Step size, 400 nm. Pixel dwell time, 2 ms. Probe power, 15 mW. IR excitation power, 0.2 mW (at 2193 cm −1 ) and 0.1 mW (at 2855 cm −1 ). Scale bars, 20 μm. (B and C) Temporal IP intensity profiles of LDs in living U2OS cells. The maximal intensities of 2193 cm −1 and 2855 cm −1 of LD1 and 2 in are plotted with time, indicated as white dotted boxes. Linear lines fitted to the temporal profiles of each IP intensity are displayed as dotted lines to show their respective trends. (D) IP spectra of the two LDs. Each data point in the IP spectra was obtained by 50 ms averaging and normalization to the intensity of the IR excitation pulse. Step size, 1 cm −1 .

Article Snippet: U2OS cell line (human bone osteosarcoma epithelial cells) and Huh-7 cell line (human hepatocellular carcinoma cells) were supplied from Korean Cell Line Bank (Seoul, Korea) and cultured in DMEM (Welgene, LM 001-17) supplemented with penicillin (100 U mL −1 ), streptomycin (100 μg mL −1 ), 10% heat-inactivated fetal bovine serum (Welgene, S1010-01) at 37 °C and pH 7.4 (5% CO 2 ).

Techniques:

Journal: Journal of Clinical Laboratory Analysis

Article Title: LncRNA LINC00210 regulated radiosensitivity of osteosarcoma cells via miR‐342‐3p/GFRA1 axis

doi: 10.1002/jcla.23540

Figure Lengend Snippet: The expression levels of LINC00210, miR‐342‐3p, and GFRA1 in osteosarcoma tissue. A, The expression of LINC00210 in osteosarcoma tissues and adjacent healthy tissues was determined by qRT‐PCR. B, The expression of miR‐342‐3p in osteosarcoma tissues and adjacent healthy tissues was determined by qRT‐PCR. C, The mRNA expression of GFRA1 in osteosarcoma tissues and adjacent healthy tissues was determined by qRT‐PCR. D, The relationship between LINC00210 and miR‐342‐3p expression was analyzed by Pearson's correlation analysis. E, The relationship between LINC00210 and GFRA1 expression was analyzed by Pearson's correlation analysis. F, Pearson's correlation analysis analyzed the relationship of miR‐342‐3p and GFRA1 expression. * P < .05

Article Snippet: Human osteoblast cell line hFOB1.19 and human osteosarcoma cell lines (SOSP‐9607 and U‐2OS) were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Clinical Laboratory Analysis

Article Title: LncRNA LINC00210 regulated radiosensitivity of osteosarcoma cells via miR‐342‐3p/GFRA1 axis

doi: 10.1002/jcla.23540

Figure Lengend Snippet: The expression levels of LINC00210, miR‐342‐3p, and GFRA1 in osteosarcoma cells. A, The expression of LINC00210 in human osteoblast cell line hFOB1.19 and osteosarcoma cell lines SOSP‐9607, U‐2OS, and F5M2 was determined by qRT‐PCR. B, The expression of miR‐342‐3p in human osteoblast cell line hFOB1.19 and osteosarcoma cell lines SOSP‐9607, U‐2OS, and F5M2 was determined by qRT‐PCR. C, The mRNA expression of GFRA1 in human osteoblast cell line hFOB1.19 and osteosarcoma cell lines SOSP‐9607, U‐2OS, and F5M2 was determined by qRT‐PCR. D, The expression of LINC00210 in SOSP‐9607 cells exposed to 4 Gy irradiation treatment was detected by qRT‐PCR. E, The expression of miR‐342‐3p in SOSP‐9607 cells exposed to 4 Gy irradiation treatment was detected by qRT‐PCR. F. The mRNA expression of GFRA1 in SOSP‐9607 cells exposed to 4 Gy irradiation treatment was detected by qRT‐PCR. * P < .05

Article Snippet: Human osteoblast cell line hFOB1.19 and human osteosarcoma cell lines (SOSP‐9607 and U‐2OS) were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Quantitative RT-PCR, Irradiation

Journal: Journal of Clinical Laboratory Analysis

Article Title: LncRNA LINC00210 regulated radiosensitivity of osteosarcoma cells via miR‐342‐3p/GFRA1 axis

doi: 10.1002/jcla.23540

Figure Lengend Snippet: Effect of LINC00210 knockdown on radiosensitivity of osteosarcoma cells. A, The expression of LINC00210 in SOSP‐9607 cells transfected with si‐LINC00210 was detected using qRT‐PCR. B, MTT assay was used to detect cell viability of SOSP‐9607 cells transfected with si‐LINC00210 and exposed to 4 Gy irradiation treatment. C, The cell apoptosis of SOSP‐9607 cells transfected with si‐LINC00210 and exposed to 4 Gy irradiation treatment was determined using Flow cytometry. D, Western blot was used to determine the levels of Cyclin D1, Bcl‐2, p21, and Bax in SOSP‐9607 cells transfected with si‐LINC00210 and exposed to 4 Gy irradiation treatment. E, The surviving fraction of SOSP‐9607 cells transfected with si‐LINC00210 and exposed to 4 Gy irradiation treatment was determined using colony formation assay. * P < .05

Article Snippet: Human osteoblast cell line hFOB1.19 and human osteosarcoma cell lines (SOSP‐9607 and U‐2OS) were obtained from American Type Culture Collection (ATCC).

Techniques: Knockdown, Expressing, Transfection, Quantitative RT-PCR, MTT Assay, Irradiation, Flow Cytometry, Western Blot, Colony Assay

Journal: Journal of Clinical Laboratory Analysis

Article Title: LncRNA LINC00210 regulated radiosensitivity of osteosarcoma cells via miR‐342‐3p/GFRA1 axis

doi: 10.1002/jcla.23540

Figure Lengend Snippet: Effect of miR‐342‐3p overexpression on radiosensitivity of osteosarcoma cells. A, The expression of miR‐342‐3p in SOSP‐9607 cells transfected with miR‐342‐3p overexpression plasmid was detected using qRT‐PCR. B, MTT assay was used to detect cell viability of SOSP‐9607 cells transfected with miR‐342‐3p overexpression plasmid and exposed to 4 Gy irradiation treatment. C, The cell apoptosis of SOSP‐9607 cells transfected with miR‐342‐3p overexpression plasmid and exposed to 4 Gy irradiation treatment was determined using Flow cytometry. D, Western blot was used to determine the levels of Cyclin D1, Bcl‐2, p21, and Bax in SOSP‐9607 cells transfected with miR‐342‐3p overexpression plasmid and exposed to 4 Gy irradiation treatment. E, The surviving fraction of SOSP‐9607 cells transfected with miR‐342‐3p overexpression plasmid and exposed to 4 Gy irradiation treatment was determined using colony formation assay. * P < .05

Article Snippet: Human osteoblast cell line hFOB1.19 and human osteosarcoma cell lines (SOSP‐9607 and U‐2OS) were obtained from American Type Culture Collection (ATCC).

Techniques: Over Expression, Expressing, Transfection, Plasmid Preparation, Quantitative RT-PCR, MTT Assay, Irradiation, Flow Cytometry, Western Blot, Colony Assay

Journal: Journal of Clinical Laboratory Analysis

Article Title: LncRNA LINC00210 regulated radiosensitivity of osteosarcoma cells via miR‐342‐3p/GFRA1 axis

doi: 10.1002/jcla.23540

Figure Lengend Snippet: LINC00210 regulated radiosensitivity of osteosarcoma cells via the miR‐342‐3p/GFRA1 axis. SOSP‐9607 cells transfected with si‐LINC00210, anti‐miR‐342‐3p, or GFRA1 plasmid and exposed to 4 Gy irradiation treatment. A, The mRNA expression of GFRA1 in SOSP‐9607 cells was determined using qRT‐PCR. B, MTT assays were used to determine the cell viability of SOSP‐9607 cells. C, Flow cytometry was used to detect cell apoptosis of SOSP‐9607 cells. D, Western blot was used to determine the levels of Cyclin D1, Bcl‐2, p21, and Bax in SOSP‐9607 cells. E, The surviving fraction of SOSP‐9607 cells was determined using colony formation assay. * P < .05

Article Snippet: Human osteoblast cell line hFOB1.19 and human osteosarcoma cell lines (SOSP‐9607 and U‐2OS) were obtained from American Type Culture Collection (ATCC).

Techniques: Transfection, Plasmid Preparation, Irradiation, Expressing, Quantitative RT-PCR, Flow Cytometry, Western Blot, Colony Assay