lamp-2a sirna  (Eurofins)

Journal: EMBO Molecular Medicine

Article Title: Chaperone-mediated autophagy regulates the metastatic state of mesenchymal tumors

doi: 10.1038/s44321-025-00210-w

Figure Lengend Snippet: Reagents and tools table

Article Snippet: siRNA targeting LAMP-2A , GenePharma (Shanghai) , 1330.

Techniques: Recombinant, Sequencing, Isolation, Plasmid Preparation, In Vivo, In Vitro, Protease Inhibitor, Bicinchoninic Acid Protein Assay, cDNA Synthesis, SYBR Green Assay, H2O2 Assay, Software, Microscopy, Inverted Microscopy, Real-time Polymerase Chain Reaction

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: List of Primer Sequences of Quantitative Real Time-PCR

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Sequencing

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: LAMP-2A expression is decreased in human lung tissue from smokers and COPD patients. ( A ) Lung homogenates from never smokers (n = 12), smokers (n = 12), and COPD patients (n = 12) were subjected to Western blot analysis for LAMP-2A and actin. ( B ) Gel data were quantified using Scion image densitometry. Data represent the mean ± SD. Normal versus smoker: ** p = 0.0194, normal versus COPD: ** p = 0.0194. ( C ) Real-time PCR analysis of LAMP2A and GAPDH expression. Data represent the mean ± SD. n.s., not-significant ( D ) LAMP-2A immunohistochemistry in lung tissues from never smokers (n = 4), smokers (n = 4), and COPD patients (n = 6). Original magnifications, ×100. ( E ) Quantity of LAMP-2A positive cells in ×100 field of the whole group. Data represent the mean ± SD. Normal versus smoker: ** p = 0.0286, normal versus COPD: ** p = 0.0095.

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Immunohistochemistry

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: Primary HBECs from smokers and COPD patients have lower expression of LAMP-2A. ( A ) Primary HBECs were isolated from normal (n = 5), smoker (n = 6), and COPD (n = 5) patients. Cell lysates were subjected to Western blot analysis for LAMP-2A and GAPDH. ( B ) Gel data were quantified using Scion image densitometry. Data represent the mean ± SD. Normal versus smoker: ** p = 0.0087, normal versus COPD: ** p = 0.0159. ( C ) Real-time PCR analysis of LAMP-2A and GAPDH expression. Data represent the mean ± SD.

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Expressing, Isolation, Western Blot, Real-time Polymerase Chain Reaction

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: LAMP-2A expression is negatively correlated with years of smoking. Pearson correlation coefficient, r, was calculated for the expression level of LAMP-2A of primary HBECs (y-axis) and clinical parameters (x-axis) ( A–D ) of 16 subjects including normal (n = 5), smokers (n = 6), and COPD patients (n = 5). ( A ) Post-bronchodilator forced expiratory volume in 1 s (FEV1% pred.) (r = 0.3593, p = 0.1718) ( B ) Diffusion capacity measured with carbon monoxide adjusted for the alveolar volume ventilated (DLCO/VA %) (r = 0.4163, p = 0.1227) ( C ) Years of smoking (r = −0.5832, ** p = 0.0177) ( D ) Aging (r = 0.2765, p = 0.2998).

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Expressing, Diffusion-based Assay

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: CSE degrades LAMP-2A via the lysosomal pathway. ( A and B ) Primary HBECs were treated with CSE (2%) for the indicated times. Total cellular extracts were subjected to Western blot analysis for LAMP-2A and GAPDH. The expression of LAMP-2A mRNA was measured by quantitative real-time PCR. Data were normalized to the expression of GAPDH. Data represent the mean ± SD. ( C ) HBECs were pretreated with z-FA-FMK (50 μM) for 1 h and then stimulated with CSE (1 or 2%) in the presence or absence of z-FA-FMK for 24 h. Total cellular extracts were subjected to Western blot analysis for LAMP-2A and GAPDH. Gel data were quantified using Scion image densitometry.

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

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

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: Downregulation of LAMP-2A is mediated by CSE-induced activation of macroautophagy. ( A and B ) Lung homogenates from never smokers (n = 12), smokers (n = 12), and COPD patients (n = 12) were subjected to Western blot analysis for LC3B and Actin ( A ). Gel data were quantified using Scion image densitometry ( B ). Data represent the mean ± SD. Normal versus smoker: p = 0.0827, normal versus COPD: ** p = 0.0447. ( C ) Primary HBECs were stimulated with CSE (2%) for the indicated times. ( D ) HBECs were treated with CSE (0.5, 1 or 2%) for 24 h. The expression of LC3B was measured by quantitative real-time PCR. Data were normalized to the expression of GAPDH. Data represent the mean ± SD. Total cell lysates were subjected to Western blot analysis for LC3B and GAPDH. ( E and F ) BEAS-2B cells were transiently transfected with control siRNA and LC3B siRNA. Forty-eight hours after transfection, the cells were stimulated with CSE (1 or 4%) for 24 h. The expression of LC3B and LAMP-2A was measured by quantitative real-time PCR. Data were normalized to the expression of GAPDH. Data represent the mean ± SD ( E ). Total cell lysates were subjected to Western blot analysis for LAMP-2A and GAPDH ( F ).

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Activation Assay, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Transfection

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: Knockdown of LAMP-2A enhanced CSE-induced cellular senescence. ( A and B ) BEAS-2B cells were transiently transfected with control siRNA and LAMP-2A siRNA. Forty-eight hours after transfection, the cells were treated with CSE (1 or 2%) for 8 or 24 h. The expression of LAMP-2A mRNA was measured by quantitative real-time PCR. Data were normalized to the expression of GAPDH. Data represent the mean ± SD. ** p < 0.0001 ( A ). Total cellular extracts were subjected to Western blot analysis for p16, p-p21, p21, p27, p-p53, p53, LAMP-2A, and GAPDH ( B ). ( C ) Whole cell lysates from primary HBECs of never smokers (n = 4), smokers (n = 4), and COPD patients (n = 4) were subjected to Western blot analysis for p16, p21, p27, and GAPDH. ( D ) Gel data were quantified using Scion image densitometry. Data represent the mean ± SD. ( E ) Lung protein extracts from never smokers (n = 8), smokers (n = 8), and COPD patients (n = 8) were subjected to Western blot analysis for p16, p21, p27, and GAPDH. ( F ) Gel data were quantified using Scion image densitometry. Data represent the mean ± SD.

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Transfection, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: Knockdown of LAMP-2A enhanced CSE-induced G2/M cell cycle arrest. ( A and B ) BEAS-2B cells were transiently transfected with control siRNA or LAMP-2A siRNA. Forty-eight hours after transfection, the cells were treated with CSE (1%) for 24 h. The cells were harvested and stained with propidium iodide (PI) for 30 min and then subjected to flow cytometric analysis to determine the cell distribution at each phase of cell cycle ( A ). Data represent the mean ± SD. Control siRNA-CSE G2/M versus LAMP-2A siRNA-CSE G2/M: ** p = 0.0351 Total cellular extracts were subjected to Western blot analysis for cyclin B1, LAMP-2A, and GAPDH ( B ).

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Transfection, Staining, Western Blot

Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Downregulation of Lysosome-Associated Membrane Protein-2A Contributes to the Pathogenesis of COPD

doi: 10.2147/COPD.S378386

Figure Lengend Snippet: Knockdown of LAMP-2A enhanced CSE-induced cell death. ( A and B ) BEAS-2B cells were transiently transfected with control siRNA or LAMP-2A siRNA. Forty-eight hours after transfection, the cells were treated with CSE (1 or 2%) for 24 h. Cell viability was determined by MTT assay ( A ). TUNEL staining was performed according to the manufacturer’s protocol ( B ). Data represent the mean ± SD. ( C ) Primary HBECs from never smokers (n = 4), smokers (n = 4), and COPD patients (n = 4) were treated with CSE (4%) for 24 h. Total cell extracts were subjected to Western blot analysis for active caspase-3, PARP, and GAPDH. ( D ) Gel data were quantified using Scion image densitometry. Data represent the mean ± SD. ( E ) TUNEL staining using human lung tissue sections of never smokers (n = 4), smokers (n = 4), and COPD patients (n = 6). Original magnifications, ×200 (left, right), ×100 (middle). ( F ) Quantity of TUNEL positive cells in ×100 field of the whole group. Data represent the mean ± SD. Normal versus smoker: ** p = 0.0294, normal versus COPD: ** p = 0.0139.

Article Snippet: Control and LAMP-2A siRNAs were purchased from Santa Cruz Biotechnology Inc. LC3B siRNA was obtained from Cell Signaling Technology.

Techniques: Transfection, MTT Assay, TUNEL Assay, Staining, Western Blot