Journal: Oncogenesis

Article Title: Neuron-specific enolase promotes stem cell-like characteristics of small-cell lung cancer by downregulating NBL1 and activating the BMP2/Smad/ID1 pathway.

doi: 10.1038/s41389-022-00396-5

Figure Lengend Snippet: Fig. 5 NSE downregulates and interacts with NBL1 in SCLC cells. A, B Western blot images showed the interaction between NSE and NBL1 by Co-IP experiment. C The double-immunofluorescence labeling experiment detected NSE and NBL1 and displayed co-location in clinical samples (original magnification ×40, scale bar 50 μm). D Western blot images show the interaction of BMP2 and BMPR1A enhanced after overexpression of NSE. E The photo shows a significant and negative correlation between ENO2 (encode NSE) and NBL1 in mRNA level using the CCLE database.

Article Snippet: The membranes were incubated with the following antibodies: NSE (1:2000, Abcam), GFP (1:2000, Proteintech), Flag (1:1000, Proteintech), OCT4 (1:1000, CST), Nanog (1:2000, CST), SOX2 (1:1000, CST), β-actin (1:5000, Proteintech), BMP2 (1:1000, Proteintech), Smad1 (1:2000, Proteintech), pSmad1/5/8 (1:1000, CST), and ID1 (1:500, Proteintech).

Techniques: Western Blot, Co-Immunoprecipitation Assay, Labeling, Over Expression

Journal: Environmental pollution (Barking, Essex : 1987)

Article Title: Biotransformation of graphene oxide within lung fluids could intensify its synergistic biotoxicity effect with cadmium by inhibiting cellular efflux of cadmium.

doi: 10.1016/j.envpol.2022.119421

Figure Lengend Snippet: Fig. 3. The cytotoxicity of Cd and intracellular Cd accumulation with or without GOs pre-treatment. (a) The cellular viability of BEAS-2B cells detected by CCK8 assay after being treated to Cd (1, 5, 10, 20, 25, 30 and 50 μM) for 24 h (n = 5, * indicate P < 0.05, compared to the untreated group). (b) The heat map of the cell viability inspected by CCK8 assay for BEAS- 2B cells after being treated to Cd (1, 5, 10, 20, 25, 30 and 50 μM) for 24 h with or without pre-treatment of P-GO, A-GO, or G-GO at the dose of 10 μg/mL, respectively (n = 5). (c) Intracellular Cd mass quan tification by ICP-MS. BEAS-2B cells were pre-treated with P-GO, A-GO, G-GO at 10 μg/mL for 24 h, and then exposed to CdCl2 at 10 μM for 24 h (n = 5), *P < 0.05. (d) Western blot analysis of MT1M and MT2A protein expression levels in BEAS-2B cells exposed to either GOs (10 μg/mL) or Cd (10 μM) or a combina tion of both (GOs + Cd) for 24 h.

Article Snippet: Antibodies (Abs) used were as follows: anti-NRF2 Ab (1:2000 dilution, Proteintech, USA), anti-SOD1 Ab (1:5000 dilution, Proteintech, USA), anti-SOD2 Ab (1:3000 dilution, Proteintech, USA), anti-MT1M (1:500 dilution, Proteintech, USA), anti-MT2A Ab (1:2000 dilution, Affinity, USA), anti-Caspase-8 Ab (1:5000 dilution, Proteintech, USA), anti-Bax Ab (1:5000 dilution, Proteintech, USA), anti-Bcl-2 Ab (1:5000 dilution, Proteintech, USA), anti-RIP1 Ab (1:2500 dilution, Proteintech, USA), anti-RIP3 Ab (1:2500 dilution, Proteintech, USA), anti-ABCB1 Ab (1:5000 dilution, Proteintech, USA), anti-ABCC1 Ab (1:2500 dilution, Proteintech, USA), anti-ABCG2 Ab (1:1000 dilution, Proteintech, USA).

Techniques: CCK-8 Assay, Western Blot, Expressing

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 3 Exosome and exosome-derived circHIF1α inhibits PK-15 cells proliferation. A–B The relative expression of circHIF1α in the precipitation or super natant of PIEC/PK-15 cells after PIEC cells treated with G. parasuis. C The relative expression of circHIF1α in PK-15 cells treated with exosomes from over expressed circHIF1α PIEC cells. D–G The proliferation of PK-15 cells treated with exosomes from PIEC cells after circHIF1α overexpression or knockdown was evaluated by CCK-8 (D and F) and EdU assay (E and G). Scale bar, 75 μm. H–I. The proliferation of PK-15 cells transfected with circHIF1α siRNA or overexpression vector was evaluated by CCK-8 (H) and EdU assay (I). Scale bar, 100 μm. J Mass spectrometric data of interaction between circHIF1α and ptotein showed the important correlated biological process. K The pathway result of mass spectrometry after RNA pulldown assay

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Derivative Assay, Expressing, Over Expression, Knockdown, CCK-8 Assay, EdU Assay, Transfection, Plasmid Preparation, Mass Spectrometry

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 4 Exosome and exosome-derived circHIF1α promotes PK-15 cells DNA damage, and mediates the G1/S phase. A–B The DNA damage level of PK-15 cells treated with exosomes from PIEC cells after circHIF1α overexpression or knockdown was detected by γ-H2AX immunofluorescence. Scale bar, 75 μm. C Cell cycle analysis was executed by flow cytometry when PK-15 cells were treated with exosomes from PIEC cells after circHIF1α knockdown. D Western blot showing the levels of DNA damage-related proteins, including γ-H2AX, NPM1, p53, and p-p53 in PK-15 cells transfected with ov-circHIF1α or ov-pLC5. E Cell cycle analysis was executed by flow cytometry after circHIF1α overexpression for 24 h. F The expression level of cell cycle-related proteins was detected in PK-15 cells after circHIF1α overexpression for 24 h by Western blot. Data are represented as mean ± SD. NS, not signifcant, *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Derivative Assay, Over Expression, Knockdown, Immunofluorescence, Cell Cycle Assay, Flow Cytometry, Western Blot, Transfection, Expressing

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 5 CircHIF1α reduces bacterial adhesion and invasion for PK-15 cell. A–C The quantity of PK-15 cells infected with G. parasuis (A)/S. aureus (B)/SS2 (C) was identified by the viable counting method. D–F PK-15 cells infected with G. parasuis(D) /S. aureus (E)/SS2 (F) was identified by IF. Scale bar, 25 μm. G–I The quantity of PK-15 cells infected with G. parasuis (G) /S. aureus (H) /SS2 (I) was identified after PK-15 cells were treated with 250nM NSC 80,467 by bacteria adhesion and invasion assays. J–L. The quantity of PK-15 cells infected with G. parasuis (J), S. aureus (K), and SS2 (L) was identified after PK-15 cells were treated with 2mM Thymidine by bacteria adhesion and invasion assays. Data are represented as mean ± SD. NS, not signifcant, *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Infection, Bacteria

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 8 Exosomal circHIF1α resists bacterial infection in vivo. A Left, Bioluminescent image showed localization of G. parasuis-exosome (20 mg/kg) or Mock-exosome (20 mg/kg) in mice after injecting through the tail vein at different times. Right, the fluorescence intensity of exosomes in vivo. n = 6 mice/ group. B The fluorescence intensity of G. parasuis-exosome (20 mg/kg) or Mock-exosome (20 mg/kg) in different organs of mice. n = 6 mice/group. C–E The effect of exosomes and circHIF1α on the survival of mice after G. parasuis (C)/SS2 (D)/S. aureus (E) infection. n = 6 mice/group. F–H The effect of exo somes and circHIF1α on G. parasuis (F)/SS2 (G)/S. aureus (H) count of different organs after mice were infected with G. parasuis. I H&E staining of various organs after mice were infected with G. parasuis followed treating with exosome or circHIF1α. Scale bar, 50 μm. n = 6 mice/group. Data are represented as mean ± SD. *p < 0.05, **p < 0.01

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Infection, In Vivo, Fluorescence, Staining

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 9 Proposed model for the potential function of exosomal circHIF1α in bacterial infection progression

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Infection

Journal: Nature Communications

Article Title: A non-canonical pathway regulates ER stress signaling and blocks ER stress-induced apoptosis and heart failure

doi: 10.1038/s41467-017-00171-w

Figure Lengend Snippet: Mechanism for AGGF1 regulation of miR-183-5p expression. a ChIP–qPCR analysis for the interaction between ZEB1 and miR-183-5p promoter DNA ( n = 4/group, ** P < 0.01). b Schematic diagram showing that the miR-183-5p promoter-luciferase reporter with two ZEB1-binding motifs. c Luciferase activity ( n = 6/group, ** P < 0.01). d Real-time RT-PCR analysis for miR-183-5p expression in H9C2 cells after AGGF1 treatment with or without ZEB1 overexpression ( n = 3/group, ** P < 0.01). e Western blot analysis for the ZEB1 level in H9C2 cells after AGGF1 treatment ( n = 5/group, * P < 0.05). f Western blot analysis for activation of ERK1/2 in H9C2 cells after AGGF1 treatment ( n = 5/group, ** P < 0.01). g Real-time RT-PCR analysis for ERK1 and ERK2 expression by ERK siRNA ( n = 3/group, ** P < 0.01). h Western blot analysis for the ZEB1 level by ERK siRNA ( n = 5/group, * P < 0.05). i Schematic model for a non-canonical ER stress signaling pathway mediated by AGGF1. Data are shown as the mean ± s.d. from at least three independent experiments. Statistical analysis was carried out by a Student’s two-tailed t -test

Article Snippet: Protein–DNA complexes were immunoprecipitated with IgG or an anti-ZEB1 antibody (2 μg; Proteintech).

Techniques: Expressing, ChIP-qPCR, Luciferase, Binding Assay, Activity Assay, Quantitative RT-PCR, Over Expression, Western Blot, Activation Assay, Two Tailed Test

Journal: Adipocyte

Article Title: Itm2a silencing rescues lamin A mediated inhibition of 3T3-L1 adipocyte differentiation.

doi: 10.1080/21623945.2017.1362510

Figure Lengend Snippet: Figure 1. LMNA inhibits differentiation of 3T3-L1 cells and increases Itm2a mRNA expression. 3T3-L1 preadipocytes were stably trans- fected with pCMV(PB)-Flag-LMNA-WT, pCMV(PB)-Flag-LMNA-R482W or empty vector (EV) control pCMV(PB) and induced to differentiate into adipocytes. (A) Adipogenesis was assessed at day 8 post induction; cells were stained for lipid droplet accumulation with Oil Red O. Oil Red O quantification was performed using ImageJ and expressed as Oil red O absorbance units (ORO a.u.). (B) Total RNA was isolated at day ¡2 of differentiation from the stably transfected 3T3-L1 preadipocytes and LMNA expression measured by qPCR using primers specific for human LMNA. (C) Immunoblot analysis of flag-LMNA WT and R482W mutant at day ¡2. (D, F) Total RNA was isolated at day 4 post induction and the expression of PPARg and CEBPa was analyzed by qPCR. (E) Immunoblot analysis of PPARg at day 4 post induc- tion. (G) Itm2a expression was analyzed at day ¡2 by qPCR. Student’s t-test (2-tailed, assuming equal variance) was used to calculate statistical significance compared with empty vector control cells, indicated as follows: D P < 0.05; D P < 0.01; D P < 0.001.

Article Snippet: Itm2a overexpression was detected with The DYKDDDDK tag Antibody (mAB, Mouse, A00187 – GenScript) and an ITM2A Rabbit Polyclonal Antibody (14407–1-AP, Proteintech).

Techniques: Expressing, Stable Transfection, Plasmid Preparation, Control, Staining, Isolation, Transfection, Western Blot, Mutagenesis

Journal: Adipocyte

Article Title: Itm2a silencing rescues lamin A mediated inhibition of 3T3-L1 adipocyte differentiation.

doi: 10.1080/21623945.2017.1362510

Figure Lengend Snippet: Figure 2. Endogenous Itm2a expression and promoter activity during 3T3-L1 differentiation (A) 3T3-L1 preadipocytes were grown to confluence (day-2) and 2 d later induction media was applied (day 0). A further 2 d later cells were supplemented with fresh media con- taining insulin (day 2). Following this, fresh media was applied every 2 d until day 8. Adipogenesis was assessed at day 8 by staining with Oil Red O. Total RNA was isolated at the indicated time points during 3T3-L1 differentiation. Itm2a and PPARg expression was ana- lyzed by qPCR. Transcript expression at the various time points is shown relative to expression at day 0. A Student’s t-test (2-tailed, assuming equal variance) was used to calculate statistical significance at day 2 in comparison to transcript levels at day 0. (B) Schematic depiction of Itm2a promoter luciferase constructs bearing a 2 kb or 0.5 kb promoter fragment cloned upstream of Gaussia luciferase in pGluc(PB) basic vector. The distance (¡1915 and ¡340) from the transcriptional start site (TSS) and previously predicted GATA and CRE binding sites are shown. (C, D) Luciferase activity (secreted) in 3T3-L1 cells stably transfected with pGluc(PB)ITM2A/2 kb and pGluc(PB) ITM2A/0.5 kb at day 1 and 2 of differentiation. Cells were induced to differentiate using full differentiation media (MDI– methylisobutyl- xanthine, dexamethasone and insulin), sub-maximal media (DI– dexamethasone and insulin) or D (dexamethasone) as indicated. Adipogenesis was assessed at day 8 by staining with Oil Red O. Luciferase activity is normalized to the pGluc(PB)basic empty vector con- trol. Student’s t-test (2-tailed, assuming equal variance) was used to calculate statistical significance compared with MDI induced cells, indicated as follows: D P < 0.05; D P < 0.01; D P < 0.001.

Article Snippet: Itm2a overexpression was detected with The DYKDDDDK tag Antibody (mAB, Mouse, A00187 – GenScript) and an ITM2A Rabbit Polyclonal Antibody (14407–1-AP, Proteintech).

Techniques: Expressing, Activity Assay, Staining, Isolation, Comparison, Luciferase, Construct, Clone Assay, Plasmid Preparation, Binding Assay, Stable Transfection, Transfection

Journal: Adipocyte

Article Title: Itm2a silencing rescues lamin A mediated inhibition of 3T3-L1 adipocyte differentiation.

doi: 10.1080/21623945.2017.1362510

Figure Lengend Snippet: Figure 3. Itm2a overexpression in 3T3-L1 differentiation. 3T3-L1 preadipocytes were stably transfected with pMSCV(PB)-Itm2a or empty vector control pMSCV(PB) plasmid and induced to differentiate into adipocytes. (A) Adipogenesis was assessed at day 8 post induction by staining with Oil Red O and quantification was performed using ImageJ and expressed as Oil red O absorbance units (ORO a.u.). (B,C) qPCR analysis of Itm2a and PPARg expression during differentiation of stably transfected 3T3-L1 cells. (D) Immunoblot analysis of PPARg at day 4 post induction. (E) Quantification of PPARg protein relative to b-actin, with mean and standard deviations determined by densi- tometry from 2 biologic replicates. A Student’s t-test (2-tailed, assuming equal variance) was used to calculate statistical significance compared with empty vector control cells, indicated as follows: D P < 0.05; D P < 0.01; D P < 0.001.

Article Snippet: Itm2a overexpression was detected with The DYKDDDDK tag Antibody (mAB, Mouse, A00187 – GenScript) and an ITM2A Rabbit Polyclonal Antibody (14407–1-AP, Proteintech).

Techniques: Over Expression, Stable Transfection, Transfection, Plasmid Preparation, Control, Staining, Expressing, Western Blot

Journal: Adipocyte

Article Title: Itm2a silencing rescues lamin A mediated inhibition of 3T3-L1 adipocyte differentiation.

doi: 10.1080/21623945.2017.1362510

Figure Lengend Snippet: Figure 4. shRNA mediated knockdown of Itm2a enhances 3T3-L1 differentiation and increases PPARg protein. 3T3-L1 preadipocytes were stably transfected with pRFP(PB).shItm2a or scramble control pRFP(PB).shControl and induced to differentiate using full induction media MDI (methylisobutylxanthine, dexamethasone and insulin), sub-maximal media DI (dexamethasone and insulin) or D (dexametha- sone) as indicated. (A) Adipogenesis was assessed at day 8 post induction by staining with Oil Red O; quantification was performed using ImageJ and expressed as Oil Red O absorbance units (ORO a.u.). (B, D) qPCR analysis of Itm2a, PPARg and CEBPa and immunoblot analysis of PPARg (E,F) expression during differentiation of 3T3-L1 cells stably transfected with pRFP(PB).shItm2a or scramble control pRFP(PB). (C) Immunoblot analysis of ITM2A knockdown in 3T3-NIH cells dual transfected with pCMV.Itm2a and pRFP(PB).shItm2a or pRFP(PB).sh.Control. (G) Quantification of PPARg protein isoforms relative to b-actin, with mean and standard deviations determined by densitometry from 2 biologic replicates. Statistical significance compared with scramble control cells indicated as follows: D P < 0.05; D P < 0.01; D P < 0.001.

Article Snippet: Itm2a overexpression was detected with The DYKDDDDK tag Antibody (mAB, Mouse, A00187 – GenScript) and an ITM2A Rabbit Polyclonal Antibody (14407–1-AP, Proteintech).

Techniques: shRNA, Knockdown, Stable Transfection, Transfection, Control, Staining, Western Blot, Expressing

Journal: Adipocyte

Article Title: Itm2a silencing rescues lamin A mediated inhibition of 3T3-L1 adipocyte differentiation.

doi: 10.1080/21623945.2017.1362510

Figure Lengend Snippet: Figure 5. shRNA mediated knockdown of Itm2a rescues LMNA inhibition of 3T3-L1 differentiation. 3T3-L1 preadipocytes were stably transfected with pCMV(PB)-Flag-LMNA-WT.shItm2a, pCMV(PB)-Flag-LMNA-WT.shControl, pCMV(PB)-Flag-LMNA-R482W.shItm2a, pCMV (PB)-Flag-LMNA-R482W.shControl or empty vector control pCMV(PB).shItm2a and pCMV(PB).shControl dual constructs. The cells were induced to differentiate. (A) Adipogenesis was assessed at day 8 post induction by staining with Oil Red O; quantification was performed using ImageJ and expressed as Oil red O absorbance units (ORO a.u.). (B) Total RNA was isolated at day ¡2 of differentiation from the stably transfected 3T3-L1 preadipocytes and LMNA expression measured by qPCR using primers specific for human LMNA. (C) Immuno- blot analysis of flag-LMNA WT and R482W mutant at day ¡2. (D) Total RNA was isolated at day 4 post induction and the expression of PPARg was analyzed by qPCR. (E) Immunoblot analysis of PPARg at day 4 post induction. Student’s t-tests (2-tailed, assuming equal var- iance) were used to calculate statistical significance compared with empty vector control cells, indicated as follows: D P < 0.05; D P < 0.01; D P < 0.001.

Article Snippet: Itm2a overexpression was detected with The DYKDDDDK tag Antibody (mAB, Mouse, A00187 – GenScript) and an ITM2A Rabbit Polyclonal Antibody (14407–1-AP, Proteintech).

Techniques: shRNA, Knockdown, Inhibition, Stable Transfection, Transfection, Plasmid Preparation, Control, Construct, Staining, Isolation, Expressing, Mutagenesis, Western Blot

Journal: Adipocyte

Article Title: Itm2a silencing rescues lamin A mediated inhibition of 3T3-L1 adipocyte differentiation.

doi: 10.1080/21623945.2017.1362510

Figure Lengend Snippet: Figure 6. Itm2a expression is downregulated in LMNA KO MEFs. (A) Total RNA was isolated from LMNA wild type (C/C) and knockout (¡/¡) MEFs and the expression of Itm2a was analyzed by qPCR. (B) Immunoblot analysis of mouse LMNA in wildtype and KO MEFs. Student’s t-test (2-tailed, assuming equal variance) was used to calculate statistical significance compared with LMNA wildtype control cells, indicated as follows: D P < 0.05; D P < 0.01; D P < 0.001.

Article Snippet: Itm2a overexpression was detected with The DYKDDDDK tag Antibody (mAB, Mouse, A00187 – GenScript) and an ITM2A Rabbit Polyclonal Antibody (14407–1-AP, Proteintech).

Techniques: Expressing, Isolation, Knock-Out, Western Blot, Control

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: A, B. Low expression of PTEN and PIK3C2A were correlated with high risk, poor prognosis and shorter OS time. C, D. High expression of ITPA and BCL3 indicated high risk, poor prognosis and shorter OS time. Kaplan-Meier survival curves were also constructed to reveal the relationship between predicted risk of ccRCC patients and the OS time. The results showed that patients with high risk had a significantly shorter OS time than those with low risk (A-D). Green and red lines indicated low- and high-risk groups, respectively. P <0.05 was considered to be statistically significant. Cens: Censored; Event: Death; Prog. Idx.: Prognosis Index.

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing, Construct

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: IHC analysis on tissue microarrays and Kaplan-Meier survival curves were constructed to verify the relationship between PTEN A. , PIK3C2A B. , ITPA C. and BCL3 D. expression with regard to OS and DFS. (A, B) Negative expression of PTEN and PIK3C2A were correlated with shorter OS and DFS time and worse prognosis. (C, D) Positive expression of ITPA and BCL3 were correlated with shorter OS and DFS time and worse prognosis. Green and blue lines indicated positive and negative expression groups, respectively. P <0.05 was considered to be statistically significant.

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Construct, Expressing

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: A. SurvExpress was used to analyze the association of the four-gene signature with the predicted risk, survival time and prognosis. B. The gene expression level of PTEN, PIK3C2A, ITPA and BCL3 were detected in high risk and low risk group. C. Kaplan-Meier survival curves showed that patients with predicted high risk ( n = 219) had significantly shorter OS time than those with low risk ( n = 249) ( P <0.05). D. ROC analysis was performed to compare the sensitivity and specificity of the survival prediction between our models. P <0.05 was considered to be statistically significant. Cens: Censored; Event: Death; Prog. Idx.: Prognosis Index; Sur.(M): Survival status (Month).

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Gene Expression

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: Correlation between clinicopathological features and the four proteins expression in ccRCC ( n =174)

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: Univariate and multivariate Cox proportional hazard regression analyses of the association of clinicopathological characteristics and the four genes' expression levels with OS

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: Univariate and multivariate Cox proportional hazard regression analyses of the association of clinicopathological characteristics and the four genes' expression levels with DFS

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing