Journal: Scientific reports

Article Title: Polyethylene glycol-phospholipid functionalized single-walled carbon nanotubes for enhanced siRNA systemic delivery.

doi: 10.1038/s41598-024-80646-1

Figure Lengend Snippet: Fig. 2. Significant gene silencing efficiency using SWNT-siRNA conjugates. (A) Graphical representation of SWNT-PL-PEG-S-S-siRNA. (B) Graphical representation of SWNT-PL-PEG-siRNA. (C, D and E) Evaluation of gene silencing effect by SWNT-PL-PEG-S-S-siGFP and SWNT-PL-PEG-siGFP conjugates in MCF-7, HEK-293T, H1299 and HeLa cells. Cells expressing stable green fluorescent protein (GFP) were treated with SWNT-PL-PEG-S-S-siGFP or SWNT-PL-PEG-siGFP conjugates at a concentration of 100 nM siGFP. The cell nuclei were stained with DAPI (blue). Microscopic observation was captured under 100x magnification. Bars represent mean ± SD of at least 3 independent experiments. * represents statistical significance as compared to vehicle controls (Student’s t-test, p < 0.01).

Article Snippet: Cell lines and cell culture The human breast cancer cell line MCF-7, human non-small cell lung carcinoma line H1299, and human cervical cancer cells HeLa were procured from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Concentration Assay, Staining

Journal: Scientific reports

Article Title: Polyethylene glycol-phospholipid functionalized single-walled carbon nanotubes for enhanced siRNA systemic delivery.

doi: 10.1038/s41598-024-80646-1

Figure Lengend Snippet: Fig. 3. Dual-silencing efficacy of siRNA-conjugated SWNT delivery system. Fluorescence microscopic images of co-expressed GFP and RFP in H1299 cells incubated with 100 nM of (A) SWNT-PL-PEG-S-S-siRNA or (B) SWNT-PL-PEG-siRNA conjugates targeting GFP (siGFP) or RFP (siRFP) or LUC (siLUC; negative control) alone, or both (dual targeting; siGFP + siRFP) for 72 h. The cell nuclei were stained with DAPI. The furthest right panel was the combined images. Microscopic observation was captured under 100x magnification.

Article Snippet: Cell lines and cell culture The human breast cancer cell line MCF-7, human non-small cell lung carcinoma line H1299, and human cervical cancer cells HeLa were procured from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Fluorescence, Incubation, Negative Control, Staining

Journal: Scientific reports

Article Title: Polyethylene glycol-phospholipid functionalized single-walled carbon nanotubes for enhanced siRNA systemic delivery.

doi: 10.1038/s41598-024-80646-1

Figure Lengend Snippet: Fig. 4. Energy dependent cellular internalization mechanisms of SWNT-siRNA conjugates independent of ABCB1. Graph shows the mean relative luminescent units (RLU) of (A and B) luciferase expressing H1299 cells co-treated with SWNT only (without siLUC), SWNT-PL-PEG-S-S-siLUC or SWNT-PL-PEG- siLUC with or without sub-lethal concentrations of various endocytosis inhibitors. NaN3, chlorpromazine, genistein and m-βCD are inhibitors for energy-dependent cellular uptake, clathrin-mediated endocytosis, caveolae-dependent endocytosis and macro-pinocytosis, respectively. Bars represent mean ± SD of at least 3 independent experiments. * represents statistical significance as compared to SWNT-siLUC (Student’s t-test, p < 0.01). (C) Western blot of ABCB1 and loading control GAPDH, cropped blots. Ectopic expression of ABCB1 (multiple drug resistance) protein in H1299-LUC cells was confirmed by the immunoblotting with increased expression of ABCB1 at 141 kDA, while GAPDH at 36 kDA served as the loading control. Full blots available in Supplementary Figure S2-S3. (D) Expression of ABCB1 did not affect the delivery of siRNA by the SWNT. Bars represent mean ± SD of at least 3 independent experiments. * represents statistical significance as compared to controls (Student’s t-test, p < 0.01).

Article Snippet: Cell lines and cell culture The human breast cancer cell line MCF-7, human non-small cell lung carcinoma line H1299, and human cervical cancer cells HeLa were procured from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Luciferase, Expressing, Western Blot, Control

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: Hsa_circ_0010235 was highly expressed in NSCLC tissues and cells. a QRT-PCR assay for the relative expression of hsa_circ_0010235 in NSCLC tissues (n = 52) and normal tissues (n = 52). b QRT-PCR assay for the relative expression of hsa_circ_0010235 in 16HBE, H1299, A549, H1581 and H23 cells. c Overall survival rate of NSCLC patients with high or low hsa_circ_0010235 expression. d , e QRT-PCR assay for the relative expression of hsa_circ_0010235 and ALDH4A1 mRNA in RNA isolated from H1299 and A549 cells digested with RNase R or not (Mock). f , g QRT-PCR assay for the half-life of hsa_circ_0010235 and ALDH4A1 in H1299 and A549 cells treated with Actinomycin D or DMSO. * P < 0.05

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing, Isolation

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: Hsa_circ_0010235 knockdown inhibited proliferation and autophagy but facilitated apoptosis in NSCLC cells. a QRT-PCR assay for the relative expression of hsa_circ_0010235 in H1299 and A549 cells transfected with si-NC, si-hsa_circ_0010235#1 or si-hsa_circ_0010235#2. b QRT-PCR assay for the relative expression of hsa_circ_0010235 in H1299 and A549 cells transfected with pCD-ciR or hsa_circ_0010235. c – j H1299 and A549 cells were transfected with si-NC, si-hsa_circ_0010235#1, pCD-ciR or hsa_circ_0010235. c, d CCK-8 assay for the cell viability of transfected cells. e Colony formation assay for the colony formation ability of transfected cells. f Flow cytometry for the apoptotic rate of transfected cells. g, h Flow cytometry for the cell cycle distribution in G0/G1, S and G2/M phases of transfected cells. i, j Western blot assay for the protein levels of LC3-I, LC3-II and p62 in transfected cells. * P < 0.05

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Knockdown, Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry, Western Blot

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: Depletion of hsa_circ_0010235 repressed migration and invasion of NSCLC cells. a – d H1299 and A549 cells were transfected with si-NC, si-hsa_circ_0010235#1, pCD-ciR or hsa_circ_0010235. a , b Transwell assay for the migration and invasion of transfected cells. c , d Wound healing assay for the migration capacity of transfected cells. * P < 0.05. The scale bar indicates 100 μm

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Migration, Transfection, Transwell Assay, Wound Healing Assay

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: Hsa_circ_0010235 served as a sponge for miR-433-3p. a Venn diagram of the overlapping target miRNAs of hsa_circ_0010235 predicted by circBank, CircInteractome and starbase. b QRT-PCR assay for the relative expression of miR-433-3p and miR-197-3p in H1299 and A549 cells transfected with pCD-ciR or hsa_circ_0010235. c The binding sites between hsa_circ_0010235 and miR-433-3p. d DLRA for the luciferase activity of H1299 and A549 cells co-transfected with miR-NC or miR-433-3p and wt-hsa_circ_0010235 or mut-hsa_circ_0010235. e RIP assay for the target relationship between hsa_circ_0010235 and miR-433-3p in H1299 and A549 cells. f QRT-PCR assay for the relative expression of miR-433-3p in 16HBE, H1299, A549, H1581 and H23 cells. g QRT-PCR assay for the relative expression of miR-433-3p in H1299 and A549 cells transfected with si-NC, si-hsa_circ_0010235#1, pCD-ciR or hsa_circ_0010235. h QRT-PCR assay for the relative expression of miR-433-3p in NSCLC tissues (n = 52) and normal tissues (n = 52). i Spearman’s correlation analysis for the expression levels of hsa_circ_0010235 and miR-433-3p in 52 NSCLC tissues. * P < 0.05

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing, Transfection, Binding Assay, Luciferase, Activity Assay

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: MiR-433-3p acted as a suppressor in NSCLC development. H1299 and A549 cells were transfected with miR-NC or miR-433-3p. a QRT-PCR assay for the relative expression of miR-433-3p in transfected cells. b , c CCK-8 assay for the cell viability of transfected cells. d Colony formation assay for the colony formation ability of transfected cells. e Flow cytometry for the apoptotic rate of transfected cells. f , g Flow cytometry for the cell cycle distribution in G0/G1, S and G2/M phases of transfected cells. h Western blot assay for the protein levels of LC3-I, LC3-II and p62 in transfected cells. i , j Transwell assay for the migration and invasion of transfected cells. k Wound healing assay for the migration capacity of transfected cells. * P < 0.05

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Transfection, Quantitative RT-PCR, Expressing, CCK-8 Assay, Colony Assay, Flow Cytometry, Western Blot, Transwell Assay, Migration, Wound Healing Assay

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: TIPRL was a target of miR-433-3p. a The binding sites between miR-433-3p and TIPRL. b DLRA for the luciferase activity of H1299 and A549 cells co-transfected with miR-NC or miR-433-3p and wt-TIPRL 3′UTR or mut-TIPRL 3′UTR. c RIP assay for the target relationship between miR-433-3p and TIPRL in H1299 and A549 cells. d , e QRT-PCR and Western blot assays for the mRNA d and protein e expression levels of TIPRL in 16HBE, H1299, A549, H1581 and H23 cells. f QRT-PCR assay for the relative mRNA expression of TIPRL in H1299 and A549 cells transfected with anti-miR-NC or anti-miR-433-3p. g , i QRT-PCR assay for the relative mRNA g and protein i expression levels of TIPRL in H1299 and A549 cells transfected with miR-NC or miR-433-3p. h , j QRT-PCR assay for the relative mRNA h and protein j expression levels of TIPRL in H1299 and A549 cells transfected with anti-miR-NC or anti-miR-433-3p. k QRT-PCR assay for the relative mRNA expression of TIPRL in NSCLC tissues (n = 52) and normal tissues (n = 52). l Western blot assay for the protein level of TIPRL in NSCLC tissues and normal tissues. m Spearman’s correlation analysis for the expression levels of miR-433-3p and TIPRL mRNA in 52 NSCLC tissues. * P < 0.05

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Quantitative RT-PCR, Western Blot, Expressing

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: Silencing of miR-433-3p or overexpression of TIPRL could attenuate hsa_circ_0010235 knockdown-induced inhibitory effects on NSCLC progression. H1299 and A549 cells were transfected with si-NC, si-hsa_circ_0010235#1, si-hsa_circ_0010235#1 + anti-miR-433-3p or hsa_circ_0010235#1 + TIPRL. a QRT-PCR assay for the relative mRNA expression of TIPRL in transfected cells. b , c Western blot assay for the protein level of TIPRL in transfected cells. d , e CCK-8 assay for the cell viability of transfected cells. f Colony formation assay for the colony formation ability of transfected cells. g Flow cytometry for the apoptotic rate of transfected cells. h , i Flow cytometry for the cell cycle distribution in G0/G1, S and G2/M phases of transfected cells. j , k Western blot assay for the protein levels of LC3-I, LC3-II and p62 in transfected cells. l , m Transwell assay for the migration and invasion of transfected cells. n Wound healing assay for the migration capacity of transfected cells. o Spearman’s correlation analysis for the expression levels of hsa_circ_0010235 and TIPRL mRNA in 52 NSCLC tissues. * P < 0.05

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Over Expression, Knockdown, Transfection, Quantitative RT-PCR, Expressing, Western Blot, CCK-8 Assay, Colony Assay, Flow Cytometry, Transwell Assay, Migration, Wound Healing Assay

Journal: Cancer Cell International

Article Title: Hsa_circ_0010235 functions as an oncogenic drive in non-small cell lung cancer by modulating miR-433-3p/TIPRL axis

doi: 10.1186/s12935-021-01764-8

Figure Lengend Snippet: Depletion of hsa_circ_0010235 blocked tumor growth in vivo. Nude mice were injected with H1299 cells stably expressing sh-hsa_circ_0010235 or sh-NC (n = 5). a Volume of formed tumors measured every 5 days. b Weight of formed tumors measured after 30 days. c , d QRT-PCR assay for the relative expression of hsa_circ_0010235 c and miR-433-3p d in formed tumors. e , f QRT-PCR and Western blot assays for the mRNA e and protein f levels of TIPRL in formed tumors. * P < 0.05

Article Snippet: Human bronchial epithelial cells 16HBE (CL-0249; Procell, Wuhan, China) and NSCLC cells H1299 (ATCC ® CRL-5803D; ATCC, Manassas, VA, USA), A549 (ATCC ® CCL-185), H1581 (ATCC ® CRL-5878) and H23 (ATCC ® CRL-5800) were cultured in Dulbecco’s Modified Eagle Medium (Gibco, Grand Island, NY, USA) mixed with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: In Vivo, Injection, Stable Transfection, Expressing, Quantitative RT-PCR, Western Blot

Journal: Computational and Structural Biotechnology Journal

Article Title: Anticancer effects and mechanisms of Pulsatilla chinensis , Bupleurum chinense and Polyporus umbellatus on human lung carcinoma and hepatoma cells

doi: 10.1016/j.csbj.2025.07.023

Figure Lengend Snippet: P. chinensis , B. chinense, and P. umbellatus showed superior anticancer effects on A549 and Huh7 cells among the 14 herbs tested.(A)Cell viability of 10 µg/ml herbal extracts treated on A549 and Huh7 cells for 48 h. (B)IC 50 of P. chinensis on A549, H1299, Huh7 and HepG2 cells. Sorafenib and Regorafenib were used as positive controls.

Article Snippet: HepG2 and H1299 cells was obtained from Cytion and HepG2.

Techniques:

Journal: Computational and Structural Biotechnology Journal

Article Title: Anticancer effects and mechanisms of Pulsatilla chinensis , Bupleurum chinense and Polyporus umbellatus on human lung carcinoma and hepatoma cells

doi: 10.1016/j.csbj.2025.07.023

Figure Lengend Snippet: P. chinensis induced similar TFs changes in A549 and Huh7 cells(A)ChEA3 enrichment reveals the top ten TFs enriched by down-regulated genes after treatment with P. chinensis in A549 (left) and Huh7 cells (right), respectively. TFs are ranked by average integrated ranks across libraries. TFs labelled with an asterisk (*) exhibit significant downregulation following P. chinensis treatment in comparison to the control group(B)IHC staining of E2F1 protein in liver and liver cancer tissue(antibody:CAB019308). All figures have been sourced from the Human Protein Atlas (HPA).(C)Western blot analysis of TFDP1 protein expression in A549, H1299, Huh7, and HepG2 cells following treatment with control, sorafenib, regorafenib, and P. chinensis. (D-E) The regulatory pathways are governed by E2F1 and TFDP, along with their downstream regulated genes in A549 (D) and Huh7 cells (E), respectively. The intensity of the colour assigned to each gene in the visualization corresponds to the log2 fold change values from the differential expression analysis. The colour blue is used to denote genes that have been downregulated.

Article Snippet: HepG2 and H1299 cells was obtained from Cytion and HepG2.

Techniques: Comparison, Control, Immunohistochemistry, Western Blot, Expressing, Quantitative Proteomics

Journal: Computational and Structural Biotechnology Journal

Article Title: Anticancer effects and mechanisms of Pulsatilla chinensis , Bupleurum chinense and Polyporus umbellatus on human lung carcinoma and hepatoma cells

doi: 10.1016/j.csbj.2025.07.023

Figure Lengend Snippet: P. chinensis as having a treatment effect similar to MDM2-p53 interaction inhibitors(A) Western blot analysis of apoptosis- and p53-related proteins in H1299, A549, Huh7, and HepG2 cells.(B) Western blot analysis of NF-κB–related proteins in Huh7 cells.(C) Bar plot illustrating MDM2 expression levels across different groups. (D) Interaction between p53 (pink) and MDM2 (grey surface representation), illustrating the native binding conformation.(E) Docking result of Nutlin-3 (green), a known p53-MDM2 inhibitor, showing its ability to occupy the key binding site and disrupt the interaction.(F) The structures of 23-hydroxybetulinic acid (23-HA). The docking result of 23-HA demonstrates its binding within the p53-MDM2 pocket, suggesting potential inhibitory activity.

Article Snippet: HepG2 and H1299 cells was obtained from Cytion and HepG2.

Techniques: Western Blot, Expressing, Binding Assay, Activity Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Therapeutic Challenge with a CDK 4/6 Inhibitor Induces an RB-dependent SMAC Mediated Apoptotic Response in Non-Small Cell Lung Cancer

doi: 10.1158/1078-0432.CCR-17-2074

Figure Lengend Snippet: (A) RB mRNA and RB immune blot analysis in RB proficient and deficient H1299 and H460 non small cell lung cancer (NSCLC) cells and the loading controls GAPDH mRNA and LaminB. (B) qRT-PCR analysis of E2F targets CyclinA and PCNA in RB proficient and deficient H1299 and H460 cells (C) Immunoblot analysis of ppRB, S780, pRB, CyclinA, PCNA and a loading control LaminB in RB proficient and deficient H1299 and H460 cells. (D) Flow cytometric analysis of BrdU incorporation in RB proficient and deficient H1299 and H460 cells in response CDK4/6 inhibitor (500 nM). (E) Graphic representation of growth analysis in RB proficient and RB deficient H460 and H1299 (shCon and shRB) cells in response to CDK4/6 inhibitor (500 nM) for 12 days in culture. (F) Graphic representation (top panel) of IVIS derived tumor luminescence of shCon and shRB H1299 cell line induced xenografts in response to CDK4/6 inhibition (150mg/kg body wt) and a graphic representation of tumor volume and tumor mass of shCon and shRB H1299 cell line induced xenografts (bottom panel) in response to CDK4/6 inhibition for three weeks (150mg/kg body wt). (G) Immunoblotting analysis of pRB, ppRBS780, LaminB, MCM7, PCNA, CDK4, CDK6, p16 and GAPDH loading control in shCon and shRB H1299 xenografts in response to CDK4/6 inhibition for three weeks (150mg/kg body wt). Each data point is a mean ± SD 5 or more animals or three or more independent experiments. **p <0.05 were considered as statistically significant.

Article Snippet: RB-proficient and deficient H1299 and H460 cells and SMAC overexpressing H1299 and H460 cells treated with either PD 0332991 or DMSO were pulse-labeled with BrdU for 2 hours prior to harvest.

Techniques: Quantitative RT-PCR, Western Blot, BrdU Incorporation Assay, Derivative Assay, Inhibition

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Therapeutic Challenge with a CDK 4/6 Inhibitor Induces an RB-dependent SMAC Mediated Apoptotic Response in Non-Small Cell Lung Cancer

doi: 10.1158/1078-0432.CCR-17-2074

Figure Lengend Snippet: (A) H&E staining of shCon and shRB H1299 and H460 cell line induced xenografts (400×) in response to CDK4/6 inhibition (150 mg/kg body wt) and a graphic representation of apoptosis positive nuclei in RB proficient and deficient H1299 and H460 cell line induced xenografts in response to CDK4/6 inhibition (left and right panel). (B) Immunoblotting analysis of cytochrome C, Cleaved caspase3 and LaminB in shCon and shRB H1299 cell line induced xenografts in response to CDK4/6 inhibition (150 mg/kg body wt) and a graphic representation of cytochrome C and cleaved caspase 3 expression in shCon and shRB tumor lysates. (C) Graphic representation of cell death analysis via photometric enzyme immunoassay in shCon and shRB H1299 cells in response to CDK4/6 inhibition (150-mg/kg body wt). (D). Immunoblotting analysis of p27, Smac Cytochrome c, cleaved caspase 3 and GAPD in shCon and shRB H460 xenograft in response to CDK4/6 inhibitor (150 mg/kg body weight). (E) Apoptotic array (protein array) images from shCon and shRB H1299 cells induced xenografts in response CDK4/6 inhibition (150-mg/kg body wt) and a graphic representation of apoptotic markers and a graphic representation of Caspase3, HTRA, p27, Smac and TNFR-1 expression in shCon and shRB tumor lysates. In each group 5 or more animals were used and each data point is a mean ± SD from 5 or more animals or three or more independent experiments. **p <0.05 were considered as statistically significant.

Article Snippet: RB-proficient and deficient H1299 and H460 cells and SMAC overexpressing H1299 and H460 cells treated with either PD 0332991 or DMSO were pulse-labeled with BrdU for 2 hours prior to harvest.

Techniques: Staining, Inhibition, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Protein Array

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Therapeutic Challenge with a CDK 4/6 Inhibitor Induces an RB-dependent SMAC Mediated Apoptotic Response in Non-Small Cell Lung Cancer

doi: 10.1158/1078-0432.CCR-17-2074

Figure Lengend Snippet: (A) Immunoblotting analysis of SMAC, cleaved caspase 3 and a loading control LaminB in RB proficient and RB deficient H1299 and H460 cells (shCon and shRB) in response to PD 0332991 (500 nM). (B) Immunoblotting analysis of SMAC, cleaved caspase 3 and a loading control LaminB in control and SMAC overexpressing H1299 and H460 cells. (C) A graphic representation of cell growth analysis in control and SMAC overexpressing H1299 and H460 cells. (D) A graphic representation of ROS (reactive oxygen species) analysis in control and SMAC overexpressing shCon and shRB H1299 and H460 cells. (E) Immunoblotting analysis of cleaved caspase 3, SMAC, PCNA, annexin V and a loading control GAPDH in SMAC proficient (shCon) and SMAC deficient (shSMAC) in H1299 cells (top panel) and a graphic representation of cell growth in SMAC proficient (shCon) and SMAC deficient (shSMAC) in H1299 cells (bottom panel) in response to PD 0332991 (500 nM). (F) A graphic representation of cell viability in SMAC proficient (shCon) and SMAC deficient (shSMAC) in H1299 cells in response to cisplatin (20 μM) or radiation (2Gy) or a combination of cisplatin and radiation. Each data point is a mean ± SD from three or more independent experiments. **p <0.05 were considered as statistically significant.

Article Snippet: RB-proficient and deficient H1299 and H460 cells and SMAC overexpressing H1299 and H460 cells treated with either PD 0332991 or DMSO were pulse-labeled with BrdU for 2 hours prior to harvest.

Techniques: Western Blot

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Therapeutic Challenge with a CDK 4/6 Inhibitor Induces an RB-dependent SMAC Mediated Apoptotic Response in Non-Small Cell Lung Cancer

doi: 10.1158/1078-0432.CCR-17-2074

Figure Lengend Snippet: (A) Gene ontology analysis in H1299 microarray data via IPA software in response to CDK4/6 inhibition for 21 days. (B) Microarray analysis derived RB/E2F signatures involved in apoptosis signaling. (C) qRT-PCR and protein validation of FOXM1, Survivin in H1299 cells in response to CDK4/6 inhibition for 21 days. (D) In silico analysis and box plot representation of RB loss signatures, FOXM1 and Survivin transcript expression in relation to RB copy number in human lung adenocarcinoma. Each data point is a mean ± SD from three or more independent experiments. **p <0.05 were considered as statistically significant.

Article Snippet: RB-proficient and deficient H1299 and H460 cells and SMAC overexpressing H1299 and H460 cells treated with either PD 0332991 or DMSO were pulse-labeled with BrdU for 2 hours prior to harvest.

Techniques: Microarray, Software, Inhibition, Derivative Assay, Quantitative RT-PCR, In Silico, Expressing

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Therapeutic Challenge with a CDK 4/6 Inhibitor Induces an RB-dependent SMAC Mediated Apoptotic Response in Non-Small Cell Lung Cancer

doi: 10.1158/1078-0432.CCR-17-2074

Figure Lengend Snippet: (A) Schematic illustration shows the location of three putative RB/E2F binding sites on FOXM1 and Survivin upstream sequence (promoters). (B & C left panels) Real-time PCR analysis of α-RB Chromatin immunoprecipitation assay shows the recruitment of RB on RB/E2F binding site II on FOXM1 and Survivin promoter, mouse α-GFP served as a negative control and input served as a positive control in shCon and shRB H1299 cells in response CDK4/6 inhibition (500 nM). (B & C right panels) PCR analysis of α-ACH4 chromatin immunoprecipitation assay shows that RB fails to recruit on RB/E2F binding site II on acetylated FOXM1 and Survivin promoter. Rabbit IgG served as a negative control and input served as a positive control in shCon and shRB cells in response to CDK4/6 inhibition (500 nM). (D) Immunoblotting analysis of FOXM1 Survivin, E2F1, E2F2, E2F3 and LaminB in ectopically expressed adenovirus harboring E2F1, E2F2, E2F3 cDNA in H1299 and H460 cells. Each data point is a mean ± SD from three or more independent experiments. **p <0.05 were considered as statistically significant.

Article Snippet: RB-proficient and deficient H1299 and H460 cells and SMAC overexpressing H1299 and H460 cells treated with either PD 0332991 or DMSO were pulse-labeled with BrdU for 2 hours prior to harvest.

Techniques: Binding Assay, Sequencing, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Negative Control, Positive Control, Inhibition, Western Blot

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Therapeutic Challenge with a CDK 4/6 Inhibitor Induces an RB-dependent SMAC Mediated Apoptotic Response in Non-Small Cell Lung Cancer

doi: 10.1158/1078-0432.CCR-17-2074

Figure Lengend Snippet: (A) Immunoblotting analysis of FOXM1, Survivin and a loading control in FOXM1 and Survivin proficient and deficient H1299 and H460 cells (left panel) and a flow cytometric analysis of BrdU incorporation in FOXM1 and Survivin proficient and deficient H1299 and H460 cells (right panel). (B) A graphic representation of cell survival analysis in FOXM1 and Survivin proficient and deficient H1299 and H460 cells (left and right panel). (C) A graphic representation of ROS (reactive oxygen species) analysis in FOXM1 and Survivin proficient and deficient H1299 and H460 cells (left and right panel). (D) Co-immunoprecipitation of FOXM1 and immune blotting analysis of FOXM1, Survivin and Caspase3 and a GAPDH loading control in shRB H1299 cells. (E) A graphic representation of cell viability analysis in FOXM1 overexpressing H1299 and H460 cells in response to PD 0332991 (left and right panel) (F) A graphic representation of cell viability analysis in Survivin overexpressing H1299 and H460 cells (left and right panel). (G) Co-immunoprecipitation of SMAC and immunoblotting analysis of FOXM1, Survivin, SMAC, Caspase 3, Cleaved Caspase3 and LaminB loading control in SMAC overexpressing shRB H1299 and H460 cells. (H) Co-immunoprecipitation of FOXM1 and immunoblotting analysis of FOXM1, Survivin and SMAC in SMAC overexpressing shRB H1299 and H460 cells. Each data point is a mean ± SD from three more independent experiments. **p <0.05 were considered as statistically significant.

Article Snippet: RB-proficient and deficient H1299 and H460 cells and SMAC overexpressing H1299 and H460 cells treated with either PD 0332991 or DMSO were pulse-labeled with BrdU for 2 hours prior to harvest.

Techniques: Western Blot, BrdU Incorporation Assay, Immunoprecipitation

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Prolonged Administration of Melatonin Ameliorates Liver Phenotypes in Cholestatic Murine Model

doi: 10.1016/j.jcmgh.2022.07.007

Figure Lengend Snippet: Patient Information

Article Snippet: , H1299 , XenoTech , Female , 17 , N/A , Caucasian , Liver.

Techniques: Control