Journal: Pharmaceutics

Article Title: Modulation of the Cytotoxic Properties of Pd(II) Complexes Based on Functionalized Carboxamides Featuring Labile Phosphoryl Coordination Sites

doi: 10.3390/pharmaceutics15041088

Figure Lengend Snippet: Cytotoxicity of the phosphoryl-functionalized amide derivatives against some human hematopoietic c ancer cell lines.

Article Snippet: To study the apoptosis inducing ability of complex 3b , K562 and K562/iS9 cells, preincubated for a day in a CO 2 incubator at 37 °C, were cultured in the medium containing 10 μM of the palladocycle for 20 h. After exposure, the cells were washed with cold PBS and incubated with Annexin V-FITC for 20 min before being treated with PI according to the supplier protocol (Elabscience Annexin V-FITC/PI Apoptosis Detection Kit).

Techniques:

Journal: Pharmaceutics

Article Title: Modulation of the Cytotoxic Properties of Pd(II) Complexes Based on Functionalized Carboxamides Featuring Labile Phosphoryl Coordination Sites

doi: 10.3390/pharmaceutics15041088

Figure Lengend Snippet: Percentages of necrotic (upper left), early apoptotic (lower right), and late apoptotic (upper right) K562 ( a , b ) and K562/iS9 ( c , d ) cells in the control experiments ( a , c ) and after exposure to complex 3b ( b , d ) for 20 h.

Article Snippet: To study the apoptosis inducing ability of complex 3b , K562 and K562/iS9 cells, preincubated for a day in a CO 2 incubator at 37 °C, were cultured in the medium containing 10 μM of the palladocycle for 20 h. After exposure, the cells were washed with cold PBS and incubated with Annexin V-FITC for 20 min before being treated with PI according to the supplier protocol (Elabscience Annexin V-FITC/PI Apoptosis Detection Kit).

Techniques: Control

Journal: Cell reports

Article Title: KIF18A promotes chromosome congression in cooperation with CENP-E downstream of CENP-C

doi: 10.1016/j.celrep.2025.116515

Figure Lengend Snippet: (A) Schematic representation of human CENP-C protein. The Mis12-binding domain (M12BD) of human CENP-C is highlighted in wild-type (WT) CENP-C. Cells expressing CENP-C lacking M12BD (CENP-C ΔM12BD cells) were generated (see also ). (B) Schematic representation of the pooled CRISPR screening in K562 WT or CENP-C ΔM12BD cells. Using CRISPR viability scores, genes which are essential for cell growth in K562 CENP-C ΔM12BD cells but not in K562 WT, cells were selected as candidates whose knockout showed synthetic lethality with CENP-C ΔM12BD . (C) Scatterplots showing the CRISPR viability score in K562 WT versus CENP-C ΔM12BD cell pools. Red dots are essential genes in K562 CENP-C ΔM12BD cells but not in K562 WT cells. (D) Gene set enrichment analysis (GSEA) for 375 candidate genes. The candidate genes were ranked by the magnitude of differences in CRISPR viability score between K562 WT and CENP-C ΔM12BD cells in descending order and applied to GSEA. The top 10 enriched gene sets were highlighted. (E) GSEA result of GOCC_SPINDLE_POLE gene set. The heatmap indicates the CRISPR viability score of the leading-edge genes of the preranking based on the magnitude of differences in CRISPR viability score between K562 WT and CENP-C ΔM12BD cells, in descending order. (F) GSEA result of GOMF_TUBULIN_BINDING gene set. The heatmap shows as in (E).

Article Snippet: To localize CENP-C, CENP-T, CENP-A, DSN1, KNL1, HEC1, and MAD2 in K562 cells, K562 cells were cytospan onto slide glasses (MATSUNAMI, S2111) or coverslips (MATSUNAMI, C024361) and then fixed with 3% paraformaldehyde (PFA; Nacalai Tesque) for 10 min at RT, and rinsed with PBS.

Techniques: Binding Assay, Expressing, Generated, CRISPR, Knock-Out

Journal: Cell reports

Article Title: KIF18A promotes chromosome congression in cooperation with CENP-E downstream of CENP-C

doi: 10.1016/j.celrep.2025.116515

Figure Lengend Snippet: (A) Cell viability of K562 WT or CENP-C ΔM12BD cells after knockout of indicated genes at 4 days after doxycycline (Dox) addition (mean and SD, two-tailed Student’s t test, CENP-C WT cells: n = 6; CENP-C ΔM12BD cells: n = 6; ** p < 0.01). (B) Representative images of DAPI-stained K562 WT or CENP-C ΔM12BD cells after knockout of indicated genes at 4 days. Scale bar, 50 μm. (C) Population of normal interphase cells, mitotic cells, and cells with micronuclei in K562 WT or CENP-C ΔM12BD cells after knockout of indicated genes at 4 days. Error bars indicate SEM. n = 3 independent experiments; 200 cells from each cell line were quantified in each experiment. (D) The growth curve of K562 WT or CENP-C ΔM12BD cells with or without KIF18A knockout. K562 WT or CENP-C ΔM12BD cells were treated with or without Dox ( KIF18A OFF or ON). The cell numbers were normalized to those at time 0 of each line. (E) Cell-cycle distribution of conditional knockout of KIF18A in K562 WT cells at each day after Dox addition, based on FACS analysis. (F) Cell-cycle distribution of conditional knockout of KIF18A in K562 CENP-C ΔM12BD cells at each day after Dox addition, based on FACS analysis. (G and H) Quantification of cells with misaligned chromosomes in K562 WT or CENP-C ΔM12BD cells with or without KIF18A knockout. The experimental scheme is shown. Cells were stained with antibodies against MAD2 (red) to detect misaligned chromosomes and CENP-T (green) as a kinetochore marker. DNA was stained with DAPI (blue). Arrowheads show typical MAD2-positive unaligned chromosomes. Scale bar, 10 μm. The cells with MAD2-positive chromosomes were quantified (H) (mean and SEM, two-tailed Student’s t test; n = 5 independent experiments; n.s., non-significant; ** p < 0.01). (I) Numbers of MAD2 positive kinetochores in each cell in each condition (WT KIF18A ON; WT KIF18A OFF; CENP-C ΔM12BD KIF18A ON; CENP-C ΔM12BD KIF18A OFF) (Mean and SEM, n = 5 independent experiments).

Article Snippet: To localize CENP-C, CENP-T, CENP-A, DSN1, KNL1, HEC1, and MAD2 in K562 cells, K562 cells were cytospan onto slide glasses (MATSUNAMI, S2111) or coverslips (MATSUNAMI, C024361) and then fixed with 3% paraformaldehyde (PFA; Nacalai Tesque) for 10 min at RT, and rinsed with PBS.

Techniques: Knock-Out, Two Tailed Test, Staining, Marker

Journal: Cell reports

Article Title: KIF18A promotes chromosome congression in cooperation with CENP-E downstream of CENP-C

doi: 10.1016/j.celrep.2025.116515

Figure Lengend Snippet: (A) Cell viability of RPE-1, K562, U2OS, A549, TIG-3, HeLa, HT1080, OVCAR-3, HT29, and HCC1806 cells with or without KIF18A knockdown (mean and SD, two-tailed Student’s t test; each sample size: n = 6; ** p < 0.01). (B–D) CENP-E (B), CENP-C (C), and CENP-T (D) localization in RPE-1, K562, U2OS, A549, TIG-3, HeLa, HT1080, OVCAR-3, HT29, and HCC1806 cells. The cell lines were treated with 50 μM monastrol for 2 h to enrich prometaphase cells. Each cell line was cytospun together with monastrol-treated RPE-1 cells expressing mScarlet-CENP-A as an internal control for immunostaining. CENP-E (green) (B), CENP-C (green) (C), and CENP-T (green) (D) were stained with antibodies against each protein. DNA was stained with DAPI (blue). CENP-T or CENP-A was stained as a kinetochore marker (red). Scale bar, 10 μm. CENP-E, CENP-C, or CENP-T signal intensities at mitotic kinetochores were quantified and normalized with those of the internal control RPE-1 mScarlet-CENP-A cells in each sample (mean and SD, two-tailed Student’s t test, each cell line: n = 10 cells; n.s., non-significant; * p < 0.1; ** p < 0.01).

Article Snippet: To localize CENP-C, CENP-T, CENP-A, DSN1, KNL1, HEC1, and MAD2 in K562 cells, K562 cells were cytospan onto slide glasses (MATSUNAMI, S2111) or coverslips (MATSUNAMI, C024361) and then fixed with 3% paraformaldehyde (PFA; Nacalai Tesque) for 10 min at RT, and rinsed with PBS.

Techniques: Knockdown, Two Tailed Test, Expressing, Control, Immunostaining, Staining, Marker

Journal: Cell reports

Article Title: KIF18A promotes chromosome congression in cooperation with CENP-E downstream of CENP-C

doi: 10.1016/j.celrep.2025.116515

Figure Lengend Snippet: In wild-type K562 and RPE-1 cells with KIF18A depletion (middle: + KIF18A KD), although KIF18A depletion enhances microtubule dynamics and increases peripheral chromosomes, CENP-E downstream of CENP-C compensates for KIF18A depletion, supporting chromosome congression and alignment, albeit with a delay in the congression. By contrast, in CENP-E-deficient cells (bottom: K562 CENP-C ΔM12BD and RPE-1 CENP-C ΔM12BD cells), KIF18A depletion disrupts chromosome congression due to weakened CENP-E activity downstream of CENP-C at kinetochores, leading to mitotic arrest and subsequent cell death. Based on these observations, we propose that the CENP-C pathway plays a role in chromosome congression, with KIF18A and CENP-E acting cooperatively to promote the congression of peripheral chromosomes during early prometaphase in wild-type K562 and RPE-1 cells (upper).

Article Snippet: To localize CENP-C, CENP-T, CENP-A, DSN1, KNL1, HEC1, and MAD2 in K562 cells, K562 cells were cytospan onto slide glasses (MATSUNAMI, S2111) or coverslips (MATSUNAMI, C024361) and then fixed with 3% paraformaldehyde (PFA; Nacalai Tesque) for 10 min at RT, and rinsed with PBS.

Techniques: Activity Assay

Journal: Non-coding RNA Research

Article Title: A novel enhancer RNA, Hmrhl, positively regulates its host gene, phkb, in chronic myelogenous leukemia

doi: 10.1016/j.ncrna.2019.08.001

Figure Lengend Snippet: Expression and coding potential analysis of Hmrhl. a. Quantitative real time PCR analysis of Hmrhl expression showed that it is expressed in all human tissues (Brain, Heart, Kidney, lung, liver, pancreas, spleen, thymus, small intestine, colon, skeletal muscle, testes, prostate, ovary, placenta, leukocyte, from left to right) examined. Lowest expression was found in skeletal muscle (SM) which was taken as control, the level of which was considered as 1 and all others were plotted in comparison to it. Highest expression was seen in spleen (spln) followed by pancreas (Pnc), testis (Tst) and other tissues. b. Northern blot detection of Hmrhl. Total RNA from HEK 293T and K562 cell lines were separated on agarose gel and subsequently hybridized with DIG labelled Hmrhl specific riboprobe to detect the transcript (i). In parallel, methylene blue staining was used to determine the size of HMRHL, using 28 S rRNA (5 kb) and 18s rRNA (1.9 kb) as reference (ii). Note that the size of Hmrhl is similar to that of 28s rRNA, revealing that Hmrhl is about 5 kb in size. c. Protein-coding potential as determined by Broad Institute's PhyloCSF data and visualized in UCSC Genome Browser, showing that Hmrhl has no coding potential. d. Circular phylogenetic tree built in iTOL (Interactive Tree of Life).

Article Snippet: Since Hmrhl locus exhibited enhancer properties in K562 Chronic Myelogenous Leukemia cells, we examined the expression profile of Hmrhl across various human cancers using a cancer specific cDNA panel (Origene, USA) by real time qPCR.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Northern Blot, Agarose Gel Electrophoresis, Staining

Journal: Non-coding RNA Research

Article Title: A novel enhancer RNA, Hmrhl, positively regulates its host gene, phkb, in chronic myelogenous leukemia

doi: 10.1016/j.ncrna.2019.08.001

Figure Lengend Snippet: Hmrhl locus exhibits hallmarks of enhancer. a. ENCODE data visualized through Integrated Genome Viewer (IGV) for DNase hypersensitive sites, p300 binding, enhancer specific histone marks, H3K27Ac and H3K4Me1 and the promoter specific histone mark, H3K4Me3 at the 5′ end of Hmrhl, only in K562 but not in GM12878 cells. Note the two prominent peaks (red) for the enhancer mark H3K27Ac in K562. b-c. Chromatin immunoprecipitation with Ab8895 (anti-H3K4Me1 antibody) and Ab4729 (anti-H3K27Ac antibody) followed by qPCR in K562 cells. Note the enrichment of both the enhancer marks at the 5′ end of Hmrhl in the IP fraction as compared to input/PIS/gene desert region (GD), that serves as a negative control.

Article Snippet: Since Hmrhl locus exhibited enhancer properties in K562 Chronic Myelogenous Leukemia cells, we examined the expression profile of Hmrhl across various human cancers using a cancer specific cDNA panel (Origene, USA) by real time qPCR.

Techniques: Binding Assay, Chromatin Immunoprecipitation, Negative Control

Journal: Non-coding RNA Research

Article Title: A novel enhancer RNA, Hmrhl, positively regulates its host gene, phkb, in chronic myelogenous leukemia

doi: 10.1016/j.ncrna.2019.08.001

Figure Lengend Snippet: Hmrhl locus exhibits hallmarks of enhancer contd. a. Encode data shows the binding of various transcription and PolII at the 5′ end of Hmrhl. We have retained the H3K27Ac peaks in this figure also for a reference. b. Schematic for chromatin interaction analysis (ChiaPET data) for Hmrhl. The large purple-black peak representing histone marks on the extreme left denotes the promoter of phkb gene while the small purple peak at the far right represents the 5'end of Hmrhl. ChiaPET data shows the interaction of Hmrhl locus with phkb promoter, as represented by two black boxes (blue arrows) connected by a black line in b. The Hmrhl locus is expanded below in c , showing that this locus has enhancer properties only in K562 cell line (orange-yellow color), but not in other cell lines like GM12878, HepG2 or hESC. Genomic segments are colour coded by ENCODE as denoted in d , with red colour signifying active promoter ( phkb promoter at far left, black arrow in b ) while orange colour represents active enhancer at Hmrhl locus at far right (red arrow in b ).

Article Snippet: Since Hmrhl locus exhibited enhancer properties in K562 Chronic Myelogenous Leukemia cells, we examined the expression profile of Hmrhl across various human cancers using a cancer specific cDNA panel (Origene, USA) by real time qPCR.

Techniques: Binding Assay

Journal: Non-coding RNA Research

Article Title: A novel enhancer RNA, Hmrhl, positively regulates its host gene, phkb, in chronic myelogenous leukemia

doi: 10.1016/j.ncrna.2019.08.001

Figure Lengend Snippet: Hmrhl is differentially expressed in various cancers. a. Expression of Hmrhl in various normal and cancer samples as observed by qPCR. Note that Hmrhl is highly upregulated in several lymphoma samples (bracket) in comparison to normal range (arrow). In fact, of all cancers, the highest levels of Hmrhl are seen in some of the lymphoma samples. b-c. qPCR analysis of Hmrhl and PHKB expression showing that both are over expressed in K562 leukemia condition as compared to GM12878 normal lymphocytes.

Article Snippet: Since Hmrhl locus exhibited enhancer properties in K562 Chronic Myelogenous Leukemia cells, we examined the expression profile of Hmrhl across various human cancers using a cancer specific cDNA panel (Origene, USA) by real time qPCR.

Techniques: Expressing

Journal: Non-coding RNA Research

Article Title: A novel enhancer RNA, Hmrhl, positively regulates its host gene, phkb, in chronic myelogenous leukemia

doi: 10.1016/j.ncrna.2019.08.001

Figure Lengend Snippet: Hmrhl functions as enhancer RNA for phkb gene. a. Lucifaerase assay showing the intense signal of reporter activity in K562 cells with insert 3 cloned in enhancer vector. Note the low level of luciferase signal obtained with insert 2 both with promoter and enhancer vectors. b. siRNA (Sigma) mediated down-regulation of Hmrhl causes down-regulation of PHKB in K562 cells treated with Hmrhl specific siRNA pool as compared to control cells without transfection and cells treated with scrambled siRNA as negative control. c-d. Smart pool siRNA (Dharmacon) were used against the Hmrhl region to downregulate Hmrhl and subsequently expression level of PHKB gene were checked by qPCR in both K562 and GM12878 cell lines. Scrambled siRNA was used as a negative control. Note the down regulation of PHKB only in K562.

Article Snippet: Since Hmrhl locus exhibited enhancer properties in K562 Chronic Myelogenous Leukemia cells, we examined the expression profile of Hmrhl across various human cancers using a cancer specific cDNA panel (Origene, USA) by real time qPCR.

Techniques: Activity Assay, Clone Assay, Plasmid Preparation, Luciferase, Transfection, Negative Control, Expressing

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: The inhibitory activity of Jac-A on tumour cells via MTT Assay (IC 50 , μ M, n = 4, mean ± SD)

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Activity Assay, MTT Assay

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A caused K562 cell apoptosis. 10 6 of K562 cells were treated with different concentrations of Jac-A for 48 h. Cells were then stained with Annexin V-FITC and propidium iodide, and analysed using flow cytometry. Four different cell populations marked as the following: live cell population (PI - AV-), early apoptosis (PI - AV+), late apoptosis (PI + AV+) and dead cells (PI + AV-). Cells were treated with no Jac-A (A) , 0.5% DMSO (B) , or 0.1, 1, 5, 10 μM/L Jac-A (C, D, E and F) . The results are one representative of three independent experiments.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Staining, Flow Cytometry

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A induced caspase-dependent apoptosis in K562 cells. (A) Cyt c release in Jac-A-induced apoptosis of K562 cells. 10 6 of K562 cells were treated with different concentration of Jac-A for 48 h. Cytosol and mitochondrial heavy membrane samples were prepared and subjected to immunoblot with anti-Cyt c specific antibodie as described in Materials and methods. The results are one representative of three independent experiments. (B) Western blot showing conspicuous cleavage of caspase-3, caspase-9, and PARP in K562 cells treated with Jac-A. K562 cells subjected to protein extract preparation after treated with 0 (control), 3, 6, 12 μM/L Jac-A for 48 h. Then, fifty micrograms extracted protein subjected to Western blot using anti-PARP, PARP, cleaved caspase-9, caspase-9, cleaved caspase-3, caspase-3, and β-actin. The results are one representative of three independent experiments. (C) Jac-A-induced apoptosis was inhibited in a concentration dependent manner by the Z-VAD-fmk. K562 cells were first treated with or without different concentrations of Z-VAD-fmk for 4 h, followed by the treatment of Jac-A (10 μM) for 48 h.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Concentration Assay, Membrane, Western Blot, Control

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Jac-A inhibited the heterodimerization of antiapoptotic proteins with pro-apoptotic proteins. Jac-A inhibited the binding between Bax with Bcl-x L or Bcl-2 (A) and Bak with Mcl-1 (B) . K562 cells were treated with indicated concentrations of Jac-A for 48 h. 150 μg of K562 cell lysates were subjected to co-immunoprecipitation using anti-Bax and anti-Bak antibodies, respectively, further immunoblot with anti- Bcl-x L , Bcl-2, Mcl-1, Bax, or Bak antibody, as described in Materials and methods. The results are one representative of three independent experiments.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Binding Assay, Immunoprecipitation, Western Blot

Journal: BMC Cancer

Article Title: Jacarelhyperol A induced apoptosis in leukaemia cancer cell through inhibition the activity of Bcl-2 proteins

doi: 10.1186/1471-2407-14-689

Figure Lengend Snippet: Therapeutic study of Jac-A in the K562-bearing mice (n = 10/group). (A) Tumour volume plot of K562-bearing mice treated with vehicle or Jac-A at 2, 10, or 50 mg/kg by oral gavage for 21 days. The tumours were measured twice per week. The data are represented as the mean ± SEM. Tumour growth was inhibited significantly after treatment with Jac-A compared with the control group. *, P < 0.05; †, P < 0.01; ‡, P < 0.001 compared with the control group. (B) Selected nude mice models of different groups treated with Jac-A or vehicle at day 14 after therapy. (C) Sizes of selected tumours harvested from dead nude mice bearing K562 cells from different groups treated with the vehicle or Jac-A. (D) Kaplan-Meier survival plot of the K562-bearing nude mice. The survival of the K562-bearing nude mice was prolonged in the Jac-A treated groups compared with control group. (E) Body weight plot of the K562-bearing nude mice. The data are represented as the mean ± SEM. *, P < 0.05; †, P < 0.01; ‡, P < 0.001 compared with the control group.

Article Snippet: 150 μg of K562 cell lysates in 500 μL of CHAPS lysis buffer were precleared for 60 min at 4°C with 20 μL of a 1:1 slurry of protein A/G Plus-Agarose (Santa Cruz Biotechnology, Cat.# sc 2003) and 1 μg of rabbit IgG.

Techniques: Control

Journal: ACS synthetic biology

Article Title: Quantitative Analyses of Core Promoters Enable Precise Engineering of Regulated Gene Expression in Mammalian Cells

doi: 10.1021/acssynbio.5b00266

Figure Lengend Snippet: Proper selection of core promoters enables restriction of antigen-stimulated T-cell activation specifically to hypoxic environments. Jurkat cells were transiently transfected with plasmids encoding a FLAG-tagged CD19 CAR expressed from a constitutive EF1α promoter or hypoxia-inducible promoters featuring either minCMV or YB_TATA as the core promoter. (a) CAR surface expression levels in transfected cells as detected by anti-FLAG antibody staining. Values shown are the means of triplicates with error bars indicating ± 1 s.d. Numbers in the plot indicate fold-induction for minCMV and YB_TATA samples. (b) Jurkat cells were cultured under normoxia for 5 hours post transfection, and then co-incubated with either parental (CD19−) or CD19+ K562 target cells for an additional 24 hours under either normoxic or hypoxic conditions. Expression of the T-cell activation marker CD69 was determined by surface antibody staining. Transfected cells were gated by dsRed+ expression prior to quantification of FLAG or CD69 staining. Data shown in (b) are representative of three independent experiments.

Article Snippet: All cells were grown in either high-glucose DMEM (MCF7 and HEK 293T; HyClone, Logan, UT) or RPMI (Jurkat and K562; Lonza, Walkersville, MA) supplemented with 10% heat inactivated FBS (HI-FBS; Life Technologies, Grand Island, NY,) at 37°C, 100% humidity and 5% CO2.

Techniques: Selection, Activation Assay, Transfection, Expressing, Staining, Cell Culture, Incubation, Marker