Journal: The Journal of Clinical Investigation

Article Title: Allosteric modulator potentiates β 2 AR agonist–promoted bronchoprotection in asthma models

doi: 10.1172/JCI167337

Figure Lengend Snippet: ( A ) HEK293 cells stably expressing the GloSensor reporter monitoring the cAMP level were pretreated with either 25 μM Cmpd-6 or the control vehicle (DMSO) and stimulated with a serial dilution of either isoproterenol (ISO) or albuterol (Alb). The level of cAMP production induced by the endogenously expressed β 2 AR was determined as described in Methods. ( B ) HEK293T cells expressing all components for the Tango assays monitoring β-arrestin recruitment to the stably expressed β 2 V 2 R as described in Methods were pretreated with Cmpd-6 or DMSO and stimulated with the agonists as described for ( A ). The extent of β-arrestin recruitment was determined as described in Methods. The values in ( A and B ) were expressed as percentage of the ISO-stimulated maximal response in the DMSO-treated condition. ( C and D ) Isolated membranes from Expi293F cells transiently expressing the human β 2 AR were incubated together with either Cmpd-6 at indicated concentrations or DMSO, a serial dilution of the indicated agonist competitor, Alb ( C ) and ISO ( D ), and 60 pM [ 125 I]-cyanopindolol ( 125 I-CYP). The reaction was terminated, and 125 I-CYP bound to the receptor was read as described in Methods. Values were normalized to the percentage of the maximal 125 I-CYP binding level obtained in each of the Cmpd-6- and DMSO-treated conditions, with data points representing mean ± SEM, obtained from 4 ( A and B ) or 5 ( C and D ) independent experiments performed in duplicate. The shift of curves was expressed as fold changes in either EC 50 and B max ( A and B ) or IC 50 (C, D) values between Cmpd-6– and DMSO-treated conditions. Statistical analyses for these shifts in each of the directions were performed using paired 2-tailed Student’s t tests. P values shown were * P < 0.05, ** P < 0.01, *** P < 0.001 compared with the control DMSO-treated condition.

Article Snippet: Human embryonic kidney-293 (HEK293) cells (ATCC) stably expressing the GloSensor cAMP reporter ( ) and HEK293T cells for the Tango assay with the β 2 V 2 R chimeric receptor ( ) were maintained at 37°C and 5% CO 2 in a humidified condition.

Techniques: Stable Transfection, Expressing, Serial Dilution, Isolation, Incubation, Binding Assay

Journal: The Journal of Clinical Investigation

Article Title: Allosteric modulator potentiates β 2 AR agonist–promoted bronchoprotection in asthma models

doi: 10.1172/JCI167337

Figure Lengend Snippet: ( A ) The sequence alignment of amino acids composing the Cmpd-6 binding site between the human and murine β 2 ARs. Shaded (yellow) amino acids represent the most essential ones, phenylalanine-133 (F-133), substituted to valine (V) in the murine receptor, and lysine-141 (K-141), for Cmpd-6 binding to the β 2 AR. ( B ) The Cmpd-6 binding site in the human (left) and modeled murine (right) β 2 ARs shows the topographical molecular surface of F-133 (purple) and V-133 (cyan) on the transmembrane-3 (TM-3). Illustrations were created with the previously reported structure PDB-6N48 using the PyMOL program. ( C ) Radioligand competition binding was performed with isolated membranes from 293ExpiF cells transiently expressing either the WT or V133F mutant murine β 2 AR as described in and D. Curve fits were plotted with data sets obtained from 4 independent experiments done in duplicate. The shift of curves was expressed as fold changes in IC 50 values between Cmpd-6– and DMSO-treated conditions. Statistical analyses for the shift in each of the WT and mutant receptor were performed using paired 2-tailed Student’s t tests. *** P adj < 0.001 compared with the DMSO-treated condition. ( D ) HEK293 cells stably expressing the GloSensor reporter were transiently transfected with one of the human WT, murine WT, and murine V133F mutant β 2 AR. After incubation with Cmpd-6 at 5 μM or DMSO vehicle, the cAMP level was monitored as described in Methods. Values were expressed as percentage of the isoproterenol-stimulated (ISO-stimulated) maximal response obtained as a control for comparable receptor expression in each transfection condition and represent mean ± SEM obtained from 4 independent experiments performed in duplicate. Statistical analyses were performed using 1-way ANOVA, repeated (related) measures with Tukey’s posthoc test. Adjusted ** P adj < 0.01.

Article Snippet: Human embryonic kidney-293 (HEK293) cells (ATCC) stably expressing the GloSensor cAMP reporter ( ) and HEK293T cells for the Tango assay with the β 2 V 2 R chimeric receptor ( ) were maintained at 37°C and 5% CO 2 in a humidified condition.

Techniques: Sequencing, Binding Assay, Isolation, Expressing, Mutagenesis, Stable Transfection, Transfection, Incubation

Journal: The Journal of Clinical Investigation

Article Title: Allosteric modulator potentiates β 2 AR agonist–promoted bronchoprotection in asthma models

doi: 10.1172/JCI167337

Figure Lengend Snippet: ( A ) The sequence alignment of amino acids composing the Cmpd-6 binding site between the human and guinea pig β 2 ARs. Shaded (yellow) amino acids represent the most essential ones, phenylalanine-133 (F-133) and lysine-141 (K-141), for Cmpd-6 binding to the β 2 AR. ( B ) Radioligand competition binding was performed with isolated membranes from 293ExpiF cells transiently expressing the guinea pig β 2 AR as described in and D. Curve fits were plotted by a 1-site competition binding-log IC 50 curve fit (GraphPad Prism) with data sets obtained from 5 independent experiments done in duplicate. The shift of curves was expressed as fold changes in IC 50 values and statistically analyzed using paired 2-tailed Student’s t test between the highest concentration of Cmpd-6– and DMSO-treated conditions. ** P < 0.01. ( C ) Cmpd-6 was incubated for 20 minutes with HEK293 cells expressing the GloSensor reporter stably and the guinea pig β 2 AR transiently. The extent of cAMP generation was determined and normalized as described for D. The value represents mean ± SEM obtained from 4 independent experiments performed in duplicate. The lines indicate the level of Cmpd-6–induced cAMP production driven by overexpression of the human (black) and the murine (red) β 2 AR shown in D. ( D – F ) Radioligand competition binding was performed as essentially described above for ( B ) with multiple β 2 agonists as competitors: fenoterol (FEN; D ), albuterol (Alb; E ), and salmeterol (Salm; F ). Curve fits, the expression of the curve shift, and statistical analyses were also generated as described for ( B ) with data sets obtained from 4 independent experiments performed in duplicate. *** P < 0.001.

Article Snippet: Human embryonic kidney-293 (HEK293) cells (ATCC) stably expressing the GloSensor cAMP reporter ( ) and HEK293T cells for the Tango assay with the β 2 V 2 R chimeric receptor ( ) were maintained at 37°C and 5% CO 2 in a humidified condition.

Techniques: Sequencing, Binding Assay, Isolation, Expressing, Concentration Assay, Incubation, Stable Transfection, Over Expression, Generated

Journal: bioRxiv

Article Title: Curcumin extends the lifespan of aging postmitotic cells with mitochondrial dysfunction

doi: 10.1101/2023.09.06.556469

Figure Lengend Snippet: The effect of curcumin treatment on the lifespan of human kidney cells (HEK293) postmitotic cells was assessed in D10 medium in a 96-well plate. The survival of aging postmitotic cells was measured using propidium iodide (PI)-fluorescent-based method for the indicated day of analysis. Data are presented as means ± SD (n=2). Statistical significance was determined as follows: *P < 0.05, ****P < 0.0001 and ns (non-significant), based on a two-way ANOVA followed by Šídák’s multiple comparisons test.

Article Snippet: Our study also involved the use of the human embryonic kidney cell line HEK293, obtained from the American Type Culture Collection (ATCC) in Manassas, Virginia, United States.

Techniques:

Journal: eLife

Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

doi: 10.7554/eLife.86976

Figure Lengend Snippet: ( A ) HEK293 cells were treated with 0.5 µM doxorubicin (DOX) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( B ) HEK293 cells were treated with 10 mM hydroxyurea (HU) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( C ) HEK293 cells were treated with 10 nM camptothecin (CPT) as indicated, cell lysates were collected and analyzed by immunoblotting for MASTL, RPA32, and α-tubulin. ( D ) HeLa cells were treated with 10 mM HU, and incubated as indicated. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, RPA32, phospho-ATM/ATR substrates, and α-tubulin. ( E ) HeLa cells were treated with or without 0.5 µM DOX, 10 nM CPT, 10 mM HU, and 20 Gy ionizing radiation (IR) for 4 hr. Cell lysates were collected and analyzed by immunoblotting for MASTL, phospho-H2AX Ser-139, phospho-ATM/ATR substrates, and α-tubulin. ( F ) HeLa cells were treated with 0.5 μM DOX, and analyzed by immunoblotting for MASTL, α-tubulin, Aurora A, Aurora B, CDK1, cyclin B1, and phospho-ATM/ATR substrates. ( G ) SCC38 cells were incubated with or without HU and caffeine, as indicated, and analyzed by immunoblotting for MASTL and β-actin.

Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

Techniques: Western Blot, Incubation

Journal: eLife

Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

doi: 10.7554/eLife.86976

Figure Lengend Snippet: ( A–C ) HeLa cells expressing CFP-MASTL were treated without or with ionizing radiation (IR). Cells were analyzed by direct fluorescence for CFP-MASTL expression, and by immunofluorescence for the phosphorylation of ATM/ATR substrates. Quantifications are shown in panels B and C (>10 cells/time point). ( D, E ) HEK293 cells were treated without ( D ) or with ( E ) 10 mM hydroxyurea (HU) for 2 hr. These cells were then treated with cycloheximide (CHX, 20 μg/ml) at time 0 to block protein synthesis, and analyzed by immunoblotting for the protein stability of MASTL and α-tubulin.

Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

Techniques: Expressing, Fluorescence, Immunofluorescence, Blocking Assay, Western Blot

Journal: eLife

Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

doi: 10.7554/eLife.86976

Figure Lengend Snippet: ( A ) The sequence alignment of the conserved E6AP Ser-218 motif in human, mouse, and Xenopus . ( B ) A phospho-specific antibody recognizing E6AP Ser-218 was generated, as described in Materials and methods. WT or E6AP knockout (KO) HEK293 cells were treated without or with 10 mM hydroxyurea (HU) for 4 hr, and analyzed by immunoblotting for phospho-E6AP Ser-218, E6AP, and α-tubulin. ( C ) HeLa cells were treated without or with 0.5 μM doxorubicin (DOX) and 5 μM KU55933 (ATMi), as indicated, for 1 hr, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-E6AP Ser-218, and α-tubulin. ( D ) HeLa cells were transfected with HA-tagged WT, S218A, or S218D E6AP. Cell lysates were harvested for immunoprecipitation (IP) assays. The input, MASTL IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL and HA. ( E ) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX for 3 hr, and harvested for IP assays. The input, HA IP, and a control IP using empty beads products were analyzed by immunoblotting for MASTL, phospho-E6AP Ser-218, and HA. ( F ) E6AP KO HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in panel D. Cells were treated with or without 0.5 μM DOX, incubated as indicated, and harvested for immunoblotting for MASTL and α-tubulin.

Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

Techniques: Sequencing, Generated, Knock-Out, Western Blot, Transfection, Immunoprecipitation, Incubation

Journal: eLife

Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

doi: 10.7554/eLife.86976

Figure Lengend Snippet: ( A ) WT or E6AP knockout HEK293 cells were treated without or with doxorubicin (DOX, 0.5 μM) for 4 hr. Cells were harvested and analyzed by immunoblotting for phospho-E6AP Ser-218 and α-tubulin. ( B ) HEK293 cells were treated without or with DOX (0.5 μM), or ATM inhibitor (KU55933, 10 μM), for 4 hr, and analyzed by immunoblotting. ( C ) HeLa cells were treated with hydroxyurea (HU, 10 mM) combined with ATM/ATR inhibitor (caffeine, 4 mM) or ATM inhibitor (KU55933, 10 μM), for 12 hr, as indicated. Cells were harvested and analyzed by immunoblotting.

Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

Techniques: Knock-Out, Western Blot

Journal: eLife

Article Title: The ATM-E6AP-MASTL axis mediates DNA damage checkpoint recovery

doi: 10.7554/eLife.86976

Figure Lengend Snippet: ( A ) E6AP knockout (KO) HeLa cells were transfected with HA-tagged WT or S218A E6AP, as in . Cells were treated with 0.1 μM etoposide (ETO) for 18 hr, and released in fresh medium for recovery. Cells were then harvested at the indicated time points (after the removal of ETO) for immunofluorescence (IF) using an anti-phospho-Aurora A/B/C antibody. The activation of Aurora phosphorylation (shown in red) and chromosome condensation (in blue) indicated mitosis. The percentages of cells in mitosis were quantified manually and shown. The mean values and standard deviations were calculated from three experiments. An unpaired two-tailed Student’s t test was used to determine the statistical significance (**p<0.01, n>500 cell numbers/measurement). ( B ) E6AP KO HeLa cells expressing HA-tagged WT or S218A E6AP, as in panel A, were treated with 2 mM hydroxyurea (HU) for 18 hr. Cells were then released in fresh medium, and incubated as indicated, for recovery. Cell cycle progression was analyzed by fluorescence-activated cell sorting (FACS). ( C ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells. Cells were treated without or with 0.1 μM ETO for 18 hr, released in fresh medium for recovery, and incubated as indicated. Cells were analyzed by immunoblotting for phospho-cyclin-dependent kinase (CDK) substrates and histone H3. ( D ) WT or S218A E6AP was expressed in E6AP KO HEK293 cells, as in panel C. Cells were treated without or with 1 μM CPT for 90 min, and analyzed by immunoblotting for phospho-ATM/ATR substrates, phospho-SMC1 Ser-957, and α-tubulin.

Article Snippet: Human cervix carcinoma (HeLa) and human embryonic kidney 293 (HEK293) cell lines were obtained and authenticated by ATCC, and maintained in Dulbecco’s modified Eagle medium (DMEM, Hyclone) with 10% fetal bovine serum (FBS, Hyclone).

Techniques: Knock-Out, Transfection, Immunofluorescence, Activation Assay, Two Tailed Test, Expressing, Incubation, Fluorescence, FACS, Western Blot

Journal: iScience

Article Title: Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation of SFTPC

doi: 10.1016/j.isci.2023.107731

Figure Lengend Snippet: High-throughput screening and identification of compounds that reduce ER stress caused by mutant SFTPC (A) Diagram illustrating the construction of XBP1-HiBiT Reporter. It was spliced under ER stress and resulted in the translation of a spliced XBP1(exon 4)-HiBiT fusion protein. The STOP sign indicates the stop codon of the open reading frame of unspliced mRNA. (B) Cellular HiBiT assay using the Nano-Glo HiBiT Lytic Detection System for HEK293 cells transfected with XBP1-HiBiT Reporter and treated with tunicamycin (0.6, 1.25, 2.5 or 5.0 μg/mL) or DMSO (0.05%) for 6 h (n = 6). Data are presented as mean ± SD. ∗∗∗p < 0.001 [One-way analysis of variant (ANOVA) with Tukey’s multiple comparisons test]. (C) Diagram illustrating the screening method. (D) Validation of high-throughput screening using the cellular HiBiT assay. HEK293 cells were co-transfected with XBP1-HiBiT Reporter and empty or L188Q-mutant SFTPC expression vector (L188Q SFTPC). n = 80 (L188Q SFTPC), n = 12 (empty vector). (E) RT-qPCR of the splicing ratio of XBP1-HiBiT Reporter (spliced/total expression) in HEK293 cells subjected to 24 h incubation with co-transfected XBP1-HiBiT Reporter and wild or L188Q SFTPC expression vectors. Data are presented as mean ± SD (n = 3 independent experiments). ∗∗p < 0.01 (Welch’s t test). (F) Scatterplot for relative luminescence units (RLU) (% of DMSO) analyzed by XBP1-HiBiT Reporter co-transfected with L188Q SFTPC expression vector in the screening of 2480 compounds. Data are presented as mean (n = 2). (G) Scatterplot showing pairwise comparisons of the result of WST-8 assay (n = 2) (% of DMSO) versus the result of HiBiT assay (from Figure 1 F) in the screening of the 2480 compounds. Red and blue characters and lines indicate each threshold to identify hit compounds. See also Figure S1 .

Article Snippet: HEK293 human embryonic kidney cell line , ATCC , CRL-1573.

Techniques: High Throughput Screening Assay, Mutagenesis, Transfection, Variant Assay, Expressing, Plasmid Preparation, Quantitative RT-PCR, Incubation

Journal: iScience

Article Title: Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation of SFTPC

doi: 10.1016/j.isci.2023.107731

Figure Lengend Snippet: Secondary screening for compounds identified in the primary screening (A) RT-qPCR of the splicing ratio of XBP1-HiBiT Reporter (spliced/total expression) in HEK293 cells subjected to 24 h incubation with co-transfected XBP1-HiBiT Reporter and wild or Y104H SFTPC expression vectors. Data are presented as mean ± SD (n = 3 independent experiments). ∗∗∗p < 0.001 (Student’s t test). (B and C) The means of Spot Intensity (B) and Total Spot Area (C) for HEK293 cells transfected with the AcGFP-wild SFTPC or AcGFP-Y104H SFTPC (n = 4, each sample is averaged over 81 field-of-view × 3 well). Data are presented as mean ± SD. ∗∗∗p < 0.001 (Student’s t test). (D) Representative microscopic images of HEK293 cells transfected with the AcGFP-wild SFTPC or AcGFP-Y104H SFTPC. Nuclei were visualized by Hoechst 33342 (blue). Scale bar = 100 μm. (E) Validation of screening using cellular HiBiT assay. HEK293 cells were co-transfected with XBP1-HiBiT Reporter and empty or Y104H-mutated SFTPC expression vector (Y104H SFTPC). n = 80 (Y104H SFTPC), n = 12 (empty vector). (F) Scatterplot showing pairwise comparisons of the result of the means of Spot Intensity (% of DMSO) for HEK293 cells transfected with the AcGFP-Y104H SFTPC (Data are presented as the mean of n = 2, each sample is averaged over 81 field-of-view) versus the means of RLU (% of DMSO) analyzed by XBP1-HiBiT Reporter co-transfected with Y104H SFTPC in the screening of 65 compounds identified in primary screening (n = 2). Red points indicate the compounds which have passed this screening. (G) Analysis of the splicing ratio of XBP1-HiBiT Reporter (spliced/total expression) by RT-qPCR in HEK293 cells subjected to 4 h incubation with co-transfected XBP1-HiBiT Reporter and Y104H SFTPC expression vectors followed by treatment with each of 9 compounds for 24 h at the indicated concentration (n = 2). Data are presented as mean ± SD. Red points indicate the compounds which have passed this screening. The Red line is FC = 0.75. See also Figure S1 .

Article Snippet: HEK293 human embryonic kidney cell line , ATCC , CRL-1573.

Techniques: Quantitative RT-PCR, Expressing, Incubation, Transfection, Plasmid Preparation, Concentration Assay

Journal: iScience

Article Title: Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation of SFTPC

doi: 10.1016/j.isci.2023.107731

Figure Lengend Snippet: CPT reduces aggregates and cell death caused by Y104H SFTPC (A and B) The means of Spot Intensity of AcGFP in each cell (A) and Total Spot Area of AcGFP in each cell (B) were analyzed for A549 cells transfected with the AcGFP-wild SFTPC or AcGFP-Y104H SFTPC for 48 h (n = 3, each sample is averaged over 81 fields of view). Data are presented as the mean ± SD. ∗∗∗p < 0.001 (Student’s t test). (C) Analysis of Spot Intensity and Total Spot Area of A549 cells transfected with the AcGFP-Y104H SFTPC for 4 h and subsequently treated with CPT (1.25, 2.5, or 5 μM) or DMSO (0.1%) for 48 h (n = 3, each sample is averaged over 81 fields of view). Data are presented as the mean ± SD. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (One-way ANOVA with Tukey’s multiple comparisons test, compared with DMSO control). (D) Representative image of A549 cells transfected with AcGFP-Y104H SFTPC for 4 h and subsequently treated with CPT (2.5 μM) or DMSO (0.1%) for 48 h. Nuclei were visualized by Hoechst 33342 (blue). Scale bar = 50 μm. (E) Representative results of WB using anti-SFTPC antibody on A549 cells transfected with the AcGFP-wild SFTPC or AcGFP-Y104H SFTPC for 4 h and subsequently treated with CPT (1.25, 2.5, or 5 μM) or DMSO (0.1%) for 48 h. α-tubulin was used as a loading control (Left panel). The signal intensities were quantified using densitometry (Right panel). Three independent experiments were conducted, and the data are presented as the mean ± SD. ∗p < 0.05 (Student’s t test, compared with DMSO control). (F) The number of double-positive cells of activated caspase-3 and SFTPC-GFP in A549 cells transfected with the AcGFP-wild SFTPC or AcGFP-Y104H SFTPC for 24 h and calculated by normalizing the number of GFP-positive cells (n = 3, each sample is averaged over 169 fields of view). Data are presented as the mean ± SD. ∗∗p < 0.01 (Student’s t test). (G) The number of double-positive cells of activated caspase-3 and SFTPC-GFP in A549 cells transfected with the AcGFP-Y104H SFTPC (Left graph) or AcGFP-wild SFTPC (Right graph) for 4 h and subsequently treated with 1.25 μM CPT or DMSO (0.1%) for 24 h, and calculated by normalizing the total cell number (n = 3, each sample is averaged over 169 fields of view). Data are presented as the mean ± SD. ∗p < 0.05 (Student’s t test). (H) The number of double-positive cells of activated caspase-3 and SFTPC-GFP in HEK293 cells transfected with the AcGFP-wild SFTPC or AcGFP-Y104H SFTPC for 24 h and calculated by normalizing the number of GFP-positive cells (n = 3, each sample is averaged over 169 fields of view). Data are presented as the mean ± SD. ∗∗∗p < 0.001 (Student’s t test). (I) The number of double-positive cells of activated caspase-3 and SFTPC-GFP in HEK293 cells transfected with the AcGFP-Y104H SFTPC (Left graph) or AcGFP-wild SFTPC (Right graph) for 4 h and subsequently treated with 2.5 μM CPT or DMSO (0.1%) for 24 h, and calculated by normalizing the total cell number (n = 3, each sample is averaged over 169 fields of view). Data are presented as the mean ± SD. ∗∗∗p < 0.001 (Student’s t test). See also Figure S3 .

Article Snippet: HEK293 human embryonic kidney cell line , ATCC , CRL-1573.

Techniques: Transfection

Journal: iScience

Article Title: Cryptotanshinone is a candidate therapeutic agent for interstitial lung disease associated with a BRICHOS-domain mutation of SFTPC

doi: 10.1016/j.isci.2023.107731

Figure Lengend Snippet:

Article Snippet: HEK293 human embryonic kidney cell line , ATCC , CRL-1573.

Techniques: Recombinant, Plasmid Preparation, Software, Cell Counting, Transfection, Membrane

Journal: Molecules

Article Title: The Phytochemical Screening and Biological Properties of Brassica napus L. var. napobrassica (Rutabaga) Seeds

doi: 10.3390/molecules28176250

Figure Lengend Snippet: Cell viability of HEK-293 after 48 h treatment with 50 µg/mL and 5 µg/mL of rutabaga seed extract. Tamoxifen was tested at 100 µM. Results are mean ± SD ( n = 3). The histograms’ different letters indicate significant difference according to Tukey’s test ( p ≤ 0.05).

Article Snippet: The anti-proliferative activity of different rutabaga seed extracts was estimated on human colon adenocarcinoma cell lines (Caco-2), and their cytotoxicity was also evaluated in human embryonic kidney cells (HEK-293) purchased both from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques:

Journal: Molecules

Article Title: The Phytochemical Screening and Biological Properties of Brassica napus L. var. napobrassica (Rutabaga) Seeds

doi: 10.3390/molecules28176250

Figure Lengend Snippet: Principal component analysis “loading plot” of the total phenolic content (TPC), the antioxidant activity (DPPH) and biological activities (anti-inflammatory activity: 15-LOX), and the cytotoxic activity (Caco-2 and HEK-293) of rutabaga seed extracts.

Article Snippet: The anti-proliferative activity of different rutabaga seed extracts was estimated on human colon adenocarcinoma cell lines (Caco-2), and their cytotoxicity was also evaluated in human embryonic kidney cells (HEK-293) purchased both from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Antioxidant Activity Assay, Activity Assay

Journal: Molecules

Article Title: The Phytochemical Screening and Biological Properties of Brassica napus L. var. napobrassica (Rutabaga) Seeds

doi: 10.3390/molecules28176250

Figure Lengend Snippet: Principal components analysis “Biplot” of the total phenolic content (TPC), the antioxidant activity (DPPH), biological activities (anti-inflammatory activity: 15-LOX), and cytotoxic activity (Caco-2 and HEK-293) of rutabaga seed extracts.

Article Snippet: The anti-proliferative activity of different rutabaga seed extracts was estimated on human colon adenocarcinoma cell lines (Caco-2), and their cytotoxicity was also evaluated in human embryonic kidney cells (HEK-293) purchased both from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Antioxidant Activity Assay, Activity Assay

Journal: iScience

Article Title: Neutron-activated, plasmonically excitable Fe-Pt-Yb 2 O 3 nanoparticles delivering anti-cancer radiation against human glioblastoma cells

doi: 10.1016/j.isci.2023.107683

Figure Lengend Snippet: Fe-Pt-Yb 2 O 3 nanoparticle clusters enter cultured cells HEK-293 cells were cultured for 24 h (A–C) without (control) and (D–F) with Fe-Pt-Yb 2 O 3 nanoparticles. (A and D) DIC images of the HEK-293 cells show the effect of 0.125 mg/mL Fe-Pt-Yb 2 O 3 nanoparticles on their viability. (B and E) Nanoparticle clusters were visualized by multi-photon microscopy, and (C and F) overlayed with the DIC images. Different size clusters of nanoparticles can be seen between and inside the cells. (G–J) After 24-h incubation, the cells were fixed, washed, and labeled with DAPI to show cell nuclei. (G) Fixed HEK-293 cells were visualized with DIC, (H) DAPI with confocal microscopy, and (I) nanoparticle clusters with multi-photon microscopy. (J) The overlay shows that nanoparticle clusters (red) can be found inside the cells.

Article Snippet: HEK-293 cells human embryonal kidney cells , ATCC , ATCC CRL-1573.

Techniques: Cell Culture, Microscopy, Incubation, Labeling, Confocal Microscopy

Journal: iScience

Article Title: Neutron-activated, plasmonically excitable Fe-Pt-Yb 2 O 3 nanoparticles delivering anti-cancer radiation against human glioblastoma cells

doi: 10.1016/j.isci.2023.107683

Figure Lengend Snippet:

Article Snippet: HEK-293 cells human embryonal kidney cells , ATCC , ATCC CRL-1573.

Techniques: Recombinant, Plasmid Preparation, Software, Microscopy