Journal: International Journal of Nanomedicine

Article Title: Docetaxel-loaded solid lipid nanoparticles suppress breast cancer cells growth with reduced myelosuppression toxicity

doi: 10.2147/IJN.S70919

Figure Lengend Snippet: Genes regulated by Taxotere ® and docetaxel-loaded solid lipid nanoparticles were confirmed by quantitative polymerase chain reaction and immunoblotting. Notes: Cell cycle-related genes of E2F8 ( A ) and OIP5 ( B ), proliferation-related genes of NASP ( C ) and SOD2 ( D ), and apoptosis-related genes of PDCD4 ( E ) and PIK3R2 ( F ) were chosen for detection by quantitative polymerase chain reaction and immunoblotting. In quantitative polymerase chain reaction detection, mock-treated cells were set as the control and samples were normalized with the control. In immunoblotting detection, β-actin was used as the loading control. Abbreviations: BSN, blank solid lipid nanoparticle; DSN, docetaxel-loaded solid lipid nanoparticle; GLU, glucose; qPCR, quantitative polymerase chain reaction; TAX, Taxotere.

Article Snippet: Primary antibodies of rabbit anti-β-actin, E2f8, MRE11A, ERBB3, IGFBP6, ATF3, CCNG2, SOD2, IGFBP3, CADM1, PDCD4, GADD45A, and MKI67 were purchased from Beijing Biosynthesis Technology Co., Ltd., (Beijing, People’s Republic of China), rabbit anti-MCM6, OIP5, and NASP were purchased from Proteintech Group, Inc., (Chicago, IL, USA), rabbit anti-FAM172A and MYB were purchased from Abgent, Inc. (San Diego, CA, USA) and rabbit anti-ATRX was purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International Journal of Nanomedicine

Article Title: Docetaxel-loaded solid lipid nanoparticles suppress breast cancer cells growth with reduced myelosuppression toxicity

doi: 10.2147/IJN.S70919

Figure Lengend Snippet: Primers used for quantitative polymerase chain reaction

Article Snippet: Primary antibodies of rabbit anti-β-actin, E2f8, MRE11A, ERBB3, IGFBP6, ATF3, CCNG2, SOD2, IGFBP3, CADM1, PDCD4, GADD45A, and MKI67 were purchased from Beijing Biosynthesis Technology Co., Ltd., (Beijing, People’s Republic of China), rabbit anti-MCM6, OIP5, and NASP were purchased from Proteintech Group, Inc., (Chicago, IL, USA), rabbit anti-FAM172A and MYB were purchased from Abgent, Inc. (San Diego, CA, USA) and rabbit anti-ATRX was purchased from GeneTex, Inc. (Irvine, CA, USA).

Techniques: Sequencing

Journal: Oncotarget

Article Title: MYCN promotes neuroblastoma malignancy by establishing a regulatory circuit with transcription factor AP4

doi: 10.18632/oncotarget.10709

Figure Lengend Snippet: Genes showing two-fold differential expression 30 hours after depletion of MYCN or TFAP4 by siRNA knockdown in BE(2)-C neuroblastoma cells

Article Snippet: TaqMan ® Assays for qPCR were: TFAP4 (Hs00231478_m1), MYCN (Hs00232074_m1); SDC1 (Hs00896423_m1); PRPS2 (Hs00267624_m1); SLC7A6 (Hs00938056_m1); CYLD (Hs00211000_m1); MFSD6 (Hs00214462_m1); FMR1 (Hs00924547_m1); C SGALNACT2 (Hs00603821_m1); DENND5B (Hs00958915_m1); SSU72 (Hs00982637_m1); LCORL (Hs00766084_m1); FAM73A (Hs01594834_m1), C19orf12 (Hs01107514_m1); OIP5-AS1 (Hs01587688_g1); COPS8 (Hs00991301_g1); HPRT (Hs02800695_m1); GUSB (Hs00939627_m1); PPIA (Hs99999904_m1). qPCR assays for TFAP4 expression using SYBR ® Green were performed using published primers [ ]. qPCR assays for EMT-associated genes using SYBR ® Green were performed using primers listed in .

Techniques: Quantitative Proteomics, Knockdown, Migration, Inhibition, De-Phosphorylation Assay, Mutagenesis, Ubiquitin Proteomics

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Opa interacting protein 5 (OIP5) expression in lung and esophageal cancers and normal tissues. (A,B) Expression of OIP5 in a normal lung tissue and 15 clinical lung cancer samples and 15 lung cancer cell lines detected by semiquantitative RT‐PCR analysis. (C,D) Expression of OIP5 in a normal esophagus and 10 clinical esophageal squamous‐cell carcinoma (ESCC) tissue samples and 10 ESCC cell lines detected by semiquantitative RT‐PCR analysis. (E) Expression of OIP5 in lung cancer cell lines, examined by western blot analyses. Expression of ACTB was served as a quantity control. (F) Subcellular localization of endogenous OIP5 protein in lung cancer SBC‐5 cells. ADC, adenocarcinoma; LCC, large cell carcinoma; SCC, small cell carcinoma.

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Opa interacting protein 5 (OIP5) expression in normal tissues and lung and esophageal cancers, and association of OIP5 expression with poorer clinical outcomes for non‐small cell lung cancer (NSCLC) and esophageal squamous‐cell carcinoma (ESCC) patients. (A) Northern blot analysis of the OIP5 transcript in 23 normal human tissues. (B) Expression of OIP5 in six normal human tissues as well as various histologic types of lung cancer and ESCC, detected by immunohistochemical staining (magnification ×100). (C) Immunohistochemical staining of OIP5 protein using anti‐OIP5 antibody in eight representative paired lung and esophageal tumors and adjacent normal lung tissues (×200). (D) Representative example of OIP5 expression in lung cancer (squamous cell carcinomas) and normal lung (top, ×100; bottom ×200). (E) Kaplan–Meier analysis of survival in NSCLC patients according to OIP5 expression level (P = 0.0053; log‐rank test). (F) Representative example of OIP5 expression in ESCC and normal esophagus (top, ×100; bottom ×200). (G) Kaplan–Meier analysis of survival in ESCC patients according to OIP5 expression level (P = 0.0168; log‐rank test). ADC, adenocarcinoma; SCC, small cell carcinoma; SCLC, small cell lung cancer.

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques: Expressing, Northern Blot, Immunohistochemical staining, Staining

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Cox’s proportional hazards model analysis of prognostic factors in patients with non‐small cell lung cancer

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques:

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Cox’s proportional hazards model analysis of prognostic factors in patients with non‐small cell lung cancer

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques:

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Association between OIP5 positivity in esophageal cancer tissues and patients’ characteristics ( n = 305)

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques: Significance Assay

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Cox’s proportional hazards model analysis of prognostic factors in patients with esophageal cancers

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques:

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Effect of Opa interacting protein 5 (OIP5) on cell growth. (A) Expression of OIP5 in response to si‐OIP5 (si‐1 and ‐2) or control siRNA (LUC and On‐Target plus/CNT) in LC319, SBC‐5 and TE2 cells, analyzed by semiquantitative RT‐PCR. (B) Viability of LC319, SBC‐5 and TE2 cells evaluated by MTT assay in response to si‐1, si‐2, si‐LUC, or si‐CNT. (C) Colony‐formation assays of LC319, SBC‐5 and TE2 cells transfected with specific siRNA for OIP5 or control siRNA. (D) Expression of OIP5 in SBC‐5 and TE9 cells examined by western blot analysis. (E,F) The cells transfected with pCAGGSn3Fc‐OIP5 or mock vector were each cultured in triplicate, and the cell viability was evaluated by the MTT assay (E) and colony‐formation assay (F).

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques: Expressing, Control, Reverse Transcription Polymerase Chain Reaction, MTT Assay, Transfection, Western Blot, Plasmid Preparation, Cell Culture, Colony Assay

Journal: Cancer Science

Article Title: Characterization of an Opa interacting protein 5 involved in lung and esophageal carcinogenesis

doi: 10.1111/j.1349-7006.2011.02167.x

Figure Lengend Snippet: Stabilization of Opa interacting protein 5 (OIP5) through its interaction with Raf1 protein. (A) Interaction of exogenous OIP5 with endogenous Raf1 protein in lung cancer SBC‐5 cells. IB, immunoblotting; IP, immunoprecipitation. (B) Expression of OIP5 and Raf1 proteins in lung cancer cell lines. (C) Effect of Raf1 knockdown on the levels of OIP5 protein, detected by semiquantitative RT‐PCR analysis and western blot analysis in SBC‐5 cells. (D) Effect of Raf1 overexpression on the levels of OIP5 protein, detected by semiquantitative RT‐PCR analysis and western blot analysis in SBC‐5 cells. ADC, adenocarcinoma; IB, immunoblotting; IP, immunoprecipitation.SCC, small cell carcinoma; SCLC, small cell lung cancer.

Article Snippet: Western blotting was performed, as previously described. ( 13 ) A commercially available rabbit polyclonal antibody to human OIP5 (Catalog No. 12142‐1‐AP, Proteintech group) was confirmed to be specific to endogenous OIP5 protein by western blot analysis using lysates of lung and esophageal cancer cell lines as well as normal airway epithelial cells.

Techniques: Western Blot, Immunoprecipitation, Expressing, Knockdown, Reverse Transcription Polymerase Chain Reaction, Over Expression

Journal: Molecular Oncology

Article Title: The Sp1/FOXC1/HOTTIP/LATS2/YAP/β‐catenin cascade promotes malignant and metastatic progression of osteosarcoma

doi: 10.1002/1878-0261.12760

Figure Lengend Snippet: FOXC1 played an essential role sustaining multiple malignant phenotypes of OS cells, both in vitro and in vivo . (A) Upon transfecting SW1353 (left panel) and 143B (right panel) cells with shFOXC1‐1, shFOXC1‐2, or control shRNA (shNC), the expression of FOXC1 protein was determined by western blot. The viability, long‐term proliferation, migration, and invasion of indicated cells at indicated time points were determined by MTT assay (B), colony‐forming assay (C), wound‐healing assay (D), and Transwell invasion assay (E), respectively, and compared between shNC and shFOXC1 cells. (F–H) shNC, shFOXC1‐1, or shFOXC1‐2 SW1353 and 143B cells were injected subcutaneously to establish xenograft tumors. The pictures of isolated xenografts after 28 days were shown in F, the growth curve in G, and the weights in H. (I–K) shNC, shFOXC1‐1, or shFOXC1‐2 SW1353 and 143B cells were injected intravenously to establish lung metastasis. The pictures of isolated lung tissues after 36 days were shown in I and microscopic metastasis detected upon HE staining shown in J and K. Error bars represent standard deviation. Student's t ‐test with four biological independent replicates was used to determine statistical significance; * P < 0.05, ** P < 0.01, *** P < 0.001, when compared to shNC cells. Scale bars: 1000 μm (D), 250 μm (E), and 100 μm (J).

Article Snippet: For HOTTIP shRNA (shHOTTIP), we designed four shRNA sequences shHOTTIP #1: 5′‐AAA AGC ATC TCA AAT TAA GCT TTG CCT CGA GGC AAA GCT TAA TTT GAG ATG C‐3′; shHOTTIP #2: 5′‐AAA AGG GAC TGA ATT CTT CGA GAT TTC TCG AGA AAT CTC AAG AAT TCA GTC CC‐3′; shHOTTIP #3: 5′‐AAA AGG TGC ACC TTA TTG ATC AAA TCT CGA GAT TTG AAT AAG GTG CAC C‐3′; shHOTTIP #4: 5′‐AAA AGC AGC CAA CAA ACT GAC TTG CCT CGA GGC AAG TCA GTT TGT TGG CTG C‐3′), cloned them into OIP5 shRNA lentiviral plasmid (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and generated lentiviral particles.

Techniques: In Vitro, In Vivo, Control, shRNA, Expressing, Western Blot, Migration, MTT Assay, Wound Healing Assay, Transwell Invasion Assay, Injection, Isolation, Staining, Standard Deviation

Journal: Molecular Oncology

Article Title: The Sp1/FOXC1/HOTTIP/LATS2/YAP/β‐catenin cascade promotes malignant and metastatic progression of osteosarcoma

doi: 10.1002/1878-0261.12760

Figure Lengend Snippet: HOTTIP recruited EZH1 and LSD1 to LATS2 promoter, enhanced promoter methylation, and suppressed the expression of LATS2. (A) The percentage distribution of HOTTIP in the nucleus and cytoplasm was examined by qRT‐PCR. GAPDH and U1 were detected as the marker for cytoplasm and nucleus, respectively. (B) EZH2 and LSD1 were identified as the potential interacting partner of HOTTIP using rpiseq software. (C) The interaction between HOTTIP and EZH2 or LSD1 was examined by RIP assay and presented as fold enrichment values relative to the amount of HOTTIP bound to IgG. (D, E) The expression of LATS2 in shHOTTIP‐1 and shHOTTIP‐2 vs. shNC cells was determined on the mRNA level by qRT‐PCR (D) and western blot (E), respectively. The levels of β‐catenin, YAP1, and p‐YAP1 were also detected by western blot (E). (F–H) SW1353 or 143B cells were transfected with siRNA targeting EZH2 (si‐EZH2‐1 and si‐EZH2‐2), LSD1 (si‐LSD1‐ and si‐LSD1‐2), or control (si‐NC). The expression levels of EZH2 (F), LSD1 (G), and LATS2 (H) were detected by qRT‐PCR. (I) The occupancy of EZH2, H3K27me3, LSD1, and H3K4me2 on LATS2 promoter in indicated cells was determined by ChIP analysis and presented as a fold value relative to the amount of IgG bound to the promoter. Error bars represent standard deviation. Student's t ‐test with three biological independent replicates was used to determine statistical significance; * P < 0.05, ** P < 0.01.

Article Snippet: For HOTTIP shRNA (shHOTTIP), we designed four shRNA sequences shHOTTIP #1: 5′‐AAA AGC ATC TCA AAT TAA GCT TTG CCT CGA GGC AAA GCT TAA TTT GAG ATG C‐3′; shHOTTIP #2: 5′‐AAA AGG GAC TGA ATT CTT CGA GAT TTC TCG AGA AAT CTC AAG AAT TCA GTC CC‐3′; shHOTTIP #3: 5′‐AAA AGG TGC ACC TTA TTG ATC AAA TCT CGA GAT TTG AAT AAG GTG CAC C‐3′; shHOTTIP #4: 5′‐AAA AGC AGC CAA CAA ACT GAC TTG CCT CGA GGC AAG TCA GTT TGT TGG CTG C‐3′), cloned them into OIP5 shRNA lentiviral plasmid (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and generated lentiviral particles.

Techniques: Methylation, Expressing, Quantitative RT-PCR, Marker, Software, Western Blot, Transfection, Control, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: Investigation of circulating lncRNAs as potential biomarkers in chronic respiratory diseases

doi: 10.1186/s12967-020-02581-9

Figure Lengend Snippet: Log 2 FC and adjusted P-values in the comparison of different groups in the replication cohort in respect of mean blood expression level of lncRNAs. Only those results are given where the adjusted P < 0.05

Article Snippet: In the replication cohort, expression level of 6 lncRNAs and 2 reference genes were detected with TaqMan Non-coding RNA Assays ( JPX : Hs0139517_g1; AC016629.8 : Hs03678951_m1; HNRNPU : Hs00402532; OIP5-AS1 : Hs01587687_g1; RP11-282O18.3 : Hs00416786_m1; RP11-325K4.3 : Hs01594146_s1) and Gene Expression Assays ( B2M : Hs99999907_m1; RPLP0 : Hs00420895_gH) with Gene Expression Master Mix (all from Applied Biosystems, Waltham, MA, USA).

Techniques: Comparison, Expressing, Control