Journal: Scientific Reports

Article Title: Correlations between TBL1XR1 and recurrence of colorectal cancer

doi: 10.1038/srep44275

Figure Lengend Snippet: ( A ) The expression level of TBL1XR1 in SW480 cells was higher than that in NCM460 cells (normal epithelial cell line), which was consistent with the RT-qPCR and IHC results. ( B ) Overexpression of TBL1XR1 in SW480 cells can increase the transcription activity of β-catenin, while TBL1XR1 knock-down showed opposite effect. ( C ) TBL1XR1 overexpression can up-regulate the protein level of Vimentin, Snail, TWIST1, and N-cadherin, while the E-cadherin expression was down-regulated. On the other hand, competitive inhibition of β-catenin can impair the EMT protein expression. MTT assay ( D ) and Transwell assay ( E ) showed that TBL1XR1 can regulate the proliferation and invasion capacities of SW480 cells through Wnt-β-catenin signaling pathway.

Article Snippet: Blots were blocked for 1 h in 5% BSA and then incubated overnight with the primary antibodies: TBL1XR1 (HPA019182, Sigma), Vimentin (10366-1-AP, Proteintech), Snai1 (13099-1-AP, Proteintech), Slug (12129-1-AP, Proteintech), TWIST1 (18125-1-AP, Proteintech), N-cadherin (13769-1-AP, Proteintech), E-cadherin (20874-1-AP, Proteintech), Cyclin D1 (60186-1-Ig, Proteintech), c-Myc (10828-1-AP, Proteintech) and β-actin (sc-1616, Santa Cruz).

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Activity Assay, Knockdown, Inhibition, MTT Assay, Transwell Assay

Journal: Scientific Reports

Article Title: Correlations between TBL1XR1 and recurrence of colorectal cancer

doi: 10.1038/srep44275

Figure Lengend Snippet: ( A ) The expression level of TBL1XR1 in SW480 cells was higher than that in NCM460 cells (normal epithelial cell line), which was consistent with the RT-qPCR and IHC results. ( B ) Overexpression of TBL1XR1 in SW480 cells can increase the transcription activity of β-catenin, while TBL1XR1 knock-down showed opposite effect. ( C ) TBL1XR1 overexpression can up-regulate the protein level of Vimentin, Snail, TWIST1, and N-cadherin, while the E-cadherin expression was down-regulated. On the other hand, competitive inhibition of β-catenin can impair the EMT protein expression. MTT assay ( D ) and Transwell assay ( E ) showed that TBL1XR1 can regulate the proliferation and invasion capacities of SW480 cells through Wnt-β-catenin signaling pathway.

Article Snippet: Blots were blocked for 1 h in 5% BSA and then incubated overnight with the primary antibodies: TBL1XR1 (HPA019182, Sigma), Vimentin (10366-1-AP, Proteintech), Snai1 (13099-1-AP, Proteintech), Slug (12129-1-AP, Proteintech), TWIST1 (18125-1-AP, Proteintech), N-cadherin (13769-1-AP, Proteintech), E-cadherin (20874-1-AP, Proteintech), Cyclin D1 (60186-1-Ig, Proteintech), c-Myc (10828-1-AP, Proteintech) and β-actin (sc-1616, Santa Cruz).

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Activity Assay, Knockdown, Inhibition, MTT Assay, Transwell Assay

Journal: International heart journal

Article Title: LIM Homeobox 2 Increases Adhesion-Regulating Molecule 1 Transcription to Facilitate the Pathological Progression of Oxidized Low-Density Lipoprotein-Stimulated Atherosclerotic Cell Models.

doi: 10.1536/ihj.22-669

Figure Lengend Snippet: Figure 3. LIM homeobox 2 (LHX2) or adhesion-regulating molecule 1 (ADRM1) knockdown alleviates endothelial–mesenchymal transition (EndMT) and the apoptosis of oxidized low-density lipoprotein (ox-LDL)-treated human umbilical vein endothelial cells (HUVECs). ox-LDL- treated HUVECs were transfected with knockdown vectors short hairpin (sh)-LHX2 or sh-ADRM1. A, B: Quantitative reverse transcrip- tion-polymerase chain reaction (qRT-PCR) and western blot to assess transfection efficiency. C: qRT-PCR and western blot to evaluate the ex- pression of EndMT-related transcription factors Snai1, Snai2, and Twist1. D: Western blot to measure the levels of the EndMT-related proteins CD31, vascular epithelium-cadherin, vimentin, and α-smooth muscle actin and the apoptotic factor cleaved caspase-3. Data were presented as mean ± standard deviation, and each experiment was repeated three times. One-way analysis of variance was applied to compare multiple groups, with Tukey’s test for post hoc multiple comparisons. *P < 0.05 versus the control or sh-negative control (NC) group; #P < 0.05 versus the NC group.

Article Snippet: Afterward, the membrane was probed for 2 hours at 37°C with diluted primary antibodies against LHX2 (ab184337, 1: 1000, Abcam, Cambridge, UK), ADRM1 (ab157218, 1: 2000, Abcam), vascular epithelium (VE)-cadherin (ab 33168, 1:1000, Abcam), cleaved caspase-3 (ab32042, 1: 1000, Abcam), CD31 (ab9498, 1:1000, Abcam), vimentin (ab92547, 1:2000, Abcam), α-smooth muscle actin (αSMA, ab108424, 1:2000, Abcam), Snai1 (13099-1-AP, 1: 500, Proteintech, Chicago, IL, USA), Snai2 (12129-1-AP, 1:1000, Proteintech), Twist1 (69366S, 1:1000, Cell Signaling Technology, Beverly, MA, USA), and GAPDH (ab 8245, 1:2000, Abcam).

Techniques: Knockdown, Transfection, Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Standard Deviation, Control, Negative Control

Journal: International heart journal

Article Title: LIM Homeobox 2 Increases Adhesion-Regulating Molecule 1 Transcription to Facilitate the Pathological Progression of Oxidized Low-Density Lipoprotein-Stimulated Atherosclerotic Cell Models.

doi: 10.1536/ihj.22-669

Figure Lengend Snippet: Figure 6. LIM homeobox 2 (LHX2) induces adhesion-regulating molecule 1 (ADRM1) transcription to accelerate endothe- lial–mesenchymal transition and apoptosis in oxidized low-density lipoprotein (ox-LDL)-treated human umbilical vein endo- thelial cells (HUVECs). Ox-LDL-treated HUVECs were transfected with short hairpin (sh)-negative control (NC) + overex- pression (oe)-NC, sh-LHX2 + oe-NC, or sh-LHX2 + oe-ADRM1. A: Cell counting kit-8 assay to examine cell viability. B: Transwell assay to assess cell invasion and migration. C: Enzyme-linked immunosorbent assay to detect tumor necrosis fac- tor-α, interleukin (IL)-1β, and IL-6 levels in cell supernatants. D: Flow cytometry to observe cell apoptosis. E: Quantitative reverse transcription-polymerase chain reaction and western blot to assess the expression of Snai1, Snai2, and Twist1. F: Western blot to evaluate the expression of CD31, vascular epithelium-cadherin, vimentin, α-smooth muscle actin, and cleaved caspase-3. Data were presented as mean ± standard deviation, and each experiment was repeated three times. One-way analy- sis of variance was applied to compare multiple groups, with Tukey’s test for post hoc multiple comparisons. *P < 0.05 versus the sh-NC + oe-NC group; #P < 0.05 versus the sh-LHX2 + oe-NC group.

Article Snippet: Afterward, the membrane was probed for 2 hours at 37°C with diluted primary antibodies against LHX2 (ab184337, 1: 1000, Abcam, Cambridge, UK), ADRM1 (ab157218, 1: 2000, Abcam), vascular epithelium (VE)-cadherin (ab 33168, 1:1000, Abcam), cleaved caspase-3 (ab32042, 1: 1000, Abcam), CD31 (ab9498, 1:1000, Abcam), vimentin (ab92547, 1:2000, Abcam), α-smooth muscle actin (αSMA, ab108424, 1:2000, Abcam), Snai1 (13099-1-AP, 1: 500, Proteintech, Chicago, IL, USA), Snai2 (12129-1-AP, 1:1000, Proteintech), Twist1 (69366S, 1:1000, Cell Signaling Technology, Beverly, MA, USA), and GAPDH (ab 8245, 1:2000, Abcam).

Techniques: Transfection, Negative Control, Cell Counting, Transwell Assay, Migration, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Reverse Transcription, Polymerase Chain Reaction, Western Blot, Expressing, Standard Deviation