Journal: BioMed research international

Article Title: Effects of Hypoxic Environment on Periodontal Tissue through the ROS/TXNIP/NLRP3 Inflammasome Pathway.

doi: 10.1155/2022/7690960

Figure Lengend Snippet: Figure 2: Effects of systemic hypoxia stimulation on the expression of TXNIP/NLRP3 signaling pathway-related factors in periodontal tissues of rats: (a) immunofluorescence staining (IF) showed that TXNIP, NLRP3, ASC, and caspase-1 in periodontal tissues. Ab: alveolar; PL: periodontal ligament; R: root. Scale bars = 25 μm. (b) RT-PCR analysis of TXNIP, NLRP3, ASC, caspase-1, and IL-1β in periodontal tissues (n = 5). (c) Western blotting analysis of TXNIP, NLRP3, ASC, caspase-1, and IL-1β in periodontal tissues (n = 5 for each group). The above data are presented as the mean ± SEM, ∗p < 0:05.

Article Snippet: For IF, the sections were incubated overnight at 4°Cwith primary antibodies (1 : 250) diluted in PBS: TXNIP (Rabbit IgG, Boster, China), NLRP3 (Rabbit IgG, Boster, China), ASC (Rabbit IgG, Immunoway, USA), and caspase-1 (Rabbit IgG, Immunoway, China) and then incubated with Alexa Fluor-488 and Alexa Fluor-555 conjugate secondary antibodies at room temperature for 1h.

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

Journal: Canadian Respiratory Journal

Article Title: LASP1 Induces Epithelial-Mesenchymal Transition in Lung Cancer through the TGF- β 1/Smad/Snail Pathway

doi: 10.1155/2021/5277409

Figure Lengend Snippet: The expressions of LASP1 in human nonmalignant adjacent tissues and lung adenocarcinoma tissues were detected by immunohistochemical staining (magnification, ×400). (a) Adjacent. (b) Adenocarcinoma.

Article Snippet: Subsequently, the tissue slides were blocked by normal goat serum and then were incubated with a 1 : 50 dilution of anti-LASP1 antibody (Boster, Wuhan, China) for 2 h at 37°C.

Techniques: Immunohistochemical staining, Staining

Journal: Canadian Respiratory Journal

Article Title: LASP1 Induces Epithelial-Mesenchymal Transition in Lung Cancer through the TGF- β 1/Smad/Snail Pathway

doi: 10.1155/2021/5277409

Figure Lengend Snippet: Average optical density values of LASP1 protein expressions by immunohistochemistry staining in human nonmalignant adjacent tissues and lung adenocarcinoma tissues ( n = 12).

Article Snippet: Subsequently, the tissue slides were blocked by normal goat serum and then were incubated with a 1 : 50 dilution of anti-LASP1 antibody (Boster, Wuhan, China) for 2 h at 37°C.

Techniques: Immunohistochemistry, Staining

Journal: Canadian Respiratory Journal

Article Title: LASP1 Induces Epithelial-Mesenchymal Transition in Lung Cancer through the TGF- β 1/Smad/Snail Pathway

doi: 10.1155/2021/5277409

Figure Lengend Snippet: (a–d) LASP1 knockdown and overexpression efficiency in TGF- β 1-treated A549 and SK-MES-1 cells. After coinfection with 2 ng/ml TGF- β 1 and lentivirus containing LASP1 shRNA or 2 ng/ml TGF- β 1 and pcDNA-LASP1, the cells were lysated to detect mRNA and protein levels of LASP1. Fold change mRNA expression levels of LASP1 were quantified by RT-qPCR in A549 (a) and SK-MES-1 (c) cells. Western blot confirmed the change of LASP1 protein levels in A549 (b) and SK-MES-1 (d) cells. All quantitative amounts were normalized against GAPDH expression and shown as mean ± SD, n = 6. ∗ p < 0.05 and ∗∗∗ p < 0.001 vs. the cell alone group without TGF- β 1 treatment, and # p < 0.05, ## p < 0.01, and ### p < 0.01 vs. the TGF- β 1-treated noninfection group.

Article Snippet: Subsequently, the tissue slides were blocked by normal goat serum and then were incubated with a 1 : 50 dilution of anti-LASP1 antibody (Boster, Wuhan, China) for 2 h at 37°C.

Techniques: Knockdown, Over Expression, shRNA, Expressing, Quantitative RT-PCR, Western Blot

Journal: Canadian Respiratory Journal

Article Title: LASP1 Induces Epithelial-Mesenchymal Transition in Lung Cancer through the TGF- β 1/Smad/Snail Pathway

doi: 10.1155/2021/5277409

Figure Lengend Snippet: (a–e) Effect of LASP1 on the pathological features of TGF- β 1-treated A549 and SK-MES-1. (a) The representative images of transwell assays showed the inhibitory or enhanced effect of LASP1 silence or overexpression under TGF- β 1-induced invasion, respectively. (b) The quantification showed the relative permeability compared to the cell alone group without TGF- β 1 treatment. (c) The representative images of wound healing assays showed the inhibitory or enhanced effect of LASP1 silence or overexpression under TGF- β 1-induced migration. (d) The quantification showed the relative migration. (e) LASP1 silence or overexpression had an inhibitory or enhanced effect on lung cancer cell's proliferation until to 72 h under TGF- β 1 treatment, respectively. Data were shown as mean ± SD, n = 3. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the cell alone group without TGF- β 1 treatment, and # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. the TGF- β 1-treated noninfection group (TGF- β 1).

Article Snippet: Subsequently, the tissue slides were blocked by normal goat serum and then were incubated with a 1 : 50 dilution of anti-LASP1 antibody (Boster, Wuhan, China) for 2 h at 37°C.

Techniques: Over Expression, Permeability, Migration

Journal: Canadian Respiratory Journal

Article Title: LASP1 Induces Epithelial-Mesenchymal Transition in Lung Cancer through the TGF- β 1/Smad/Snail Pathway

doi: 10.1155/2021/5277409

Figure Lengend Snippet: (a-b) LASP1 regulated EMT-related markers expression in A549 (a) and SK-MES-1 (b) cell lines under TGF- β 1 treatment. After 48 h coinfection with TGF- β 1 accompanying LASP1 knockdown or overexpression, cell lysates were detected by western blot for EMT-related marker expression, including N-cadherin, vimentin, and E-cadherin. GAPDH was used as reference control. Data were shown as mean ± SD, n = 3. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the cell alone group without TGF- β 1 treatment, and # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. the TGF- β 1-treated noninfection group (TGF- β 1).

Article Snippet: Subsequently, the tissue slides were blocked by normal goat serum and then were incubated with a 1 : 50 dilution of anti-LASP1 antibody (Boster, Wuhan, China) for 2 h at 37°C.

Techniques: Expressing, Knockdown, Over Expression, Western Blot, Marker, Control

Journal: Canadian Respiratory Journal

Article Title: LASP1 Induces Epithelial-Mesenchymal Transition in Lung Cancer through the TGF- β 1/Smad/Snail Pathway

doi: 10.1155/2021/5277409

Figure Lengend Snippet: (a-b) LASP1 effect on key molecules in the TGF- β 1/Smad/Snail signaling pathway in A549 (a) and SK-MES-1 (b) cell lines under TGF- β 1 treatment. After 48 h coinfection with TGF- β 1 accompanying LASP1 knockdown or overexpression, the cell lysates were detected for protein levels of pSmad2, Smad2, pSmad3, Smad3, and Smad7 by western blot. Ratios of pSmad2/Smad2, pSmad3/Smad3 and Smad7/GAPDH were presented by gray intensity analysis. GAPDH was used as reference control. Data were shown as mean ± SD, n = 3. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 vs. the cell alone group without TGF- β 1 treatment, and # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. the TGF- β 1-treated noninfection group (TGF- β 1).

Article Snippet: Subsequently, the tissue slides were blocked by normal goat serum and then were incubated with a 1 : 50 dilution of anti-LASP1 antibody (Boster, Wuhan, China) for 2 h at 37°C.

Techniques: Knockdown, Over Expression, Western Blot, Control

Journal: Canadian Respiratory Journal

Article Title: LASP1 Induces Epithelial-Mesenchymal Transition in Lung Cancer through the TGF- β 1/Smad/Snail Pathway

doi: 10.1155/2021/5277409

Figure Lengend Snippet: (a–c) LASP1 effect on nuclear levels of key molecules in the TGF- β 1/Smad/Snail signaling pathway in A549 and SK-MES-1 cell lines under TGF- β 1 treatment. After 48 h coinfection with TGF- β 1 accompanying LASP1 knockdown or overexpression, the nuclear fractions were detected for protein levels of pSmad2, pSmad3, and Snail1 by western blot. (a) α -Tubulin (cytoplasmic marker) and Lamin B (nuclear marker) demonstrated the purified nuclear extracts. The representative images of western blot and the quantification normalized with (b) Lamin B in A549 and (c) SK-MES-1 cells. Data were shown as mean ± SD, n = 3. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 vs . the cell alone group without TGF- β 1 treatment, and # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. the TGF- β 1-treated noninfection group (TGF- β 1).

Article Snippet: Subsequently, the tissue slides were blocked by normal goat serum and then were incubated with a 1 : 50 dilution of anti-LASP1 antibody (Boster, Wuhan, China) for 2 h at 37°C.

Techniques: Knockdown, Over Expression, Western Blot, Marker, Purification