shrnas against lamp2a  (Genechem)

Journal: PLOS One

Article Title: LAMP2A-dependent chaperone-mediated autophagy enhances oxidative stress resistance in gastric cancer cells through selective degradation of accumulated oxidized DJ-1

doi: 10.1371/journal.pone.0331823

Figure Lengend Snippet: (A) Immunohistochemical staining of LAMP2A in gastric tumor and normal tissue. (Scale bar: 20 μm). (B) The TCGA and GEO databases show the mRNA expression levels of LAMP2A in gastric tumors and normal tissue. (C)The qPCR results showed the LAMP2A expression at mRNA levels. (D) Western blot analysis of the level of LAMP2A in several cell lines. Note:TCGA( https://portal.gdc.cancer.gov/)GEO(https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE63089 ). Data are presented as mean±SD of three independent experiments. ns, no significance, *P < 0.05. **P < 0.01. ***P < 0.001.

Article Snippet: The lentiviral vectors with shRNAs against LAMP2A were purchased from GeneChem Company, Shanghai, China (PIEL248064052).

Techniques: Immunohistochemical staining, Staining, Expressing, Western Blot

Journal: PLOS One

Article Title: LAMP2A-dependent chaperone-mediated autophagy enhances oxidative stress resistance in gastric cancer cells through selective degradation of accumulated oxidized DJ-1

doi: 10.1371/journal.pone.0331823

Figure Lengend Snippet: Four cell lines were exposed to H 2 O 2 (150μM, 300μM), respectively, to establish a severe oxidative stress cell model. WB results showed the protein expression of LAMP2A. Data are presented as mean±SD of three independent experiments. ns, no significance, *P < 0.05. **P < 0.01.

Article Snippet: The lentiviral vectors with shRNAs against LAMP2A were purchased from GeneChem Company, Shanghai, China (PIEL248064052).

Techniques: Expressing

Journal: PLOS One

Article Title: LAMP2A-dependent chaperone-mediated autophagy enhances oxidative stress resistance in gastric cancer cells through selective degradation of accumulated oxidized DJ-1

doi: 10.1371/journal.pone.0331823

Figure Lengend Snippet: (A, B) Verification of LAMP2A knockdown efficiency in MKN45 cell. (C, D) Verification of LAMP2A overexpression efficiency in AGS cell. Note: NC, negative control. Data are presented as mean±SD of three independent experiments. ns, no significance, *P < 0.05. **P < 0.01.

Article Snippet: The lentiviral vectors with shRNAs against LAMP2A were purchased from GeneChem Company, Shanghai, China (PIEL248064052).

Techniques: Knockdown, Over Expression, Negative Control

Journal: PLOS One

Article Title: LAMP2A-dependent chaperone-mediated autophagy enhances oxidative stress resistance in gastric cancer cells through selective degradation of accumulated oxidized DJ-1

doi: 10.1371/journal.pone.0331823

Figure Lengend Snippet: (A): Proliferation of the control group and LAMP2A-knockdown group. (B): Proliferation of the control group and LAMP2A-knockdown group following 150 μM H 2 O 2 treatment. (C): Proliferation of the control group and LAMP2A-overexpress group. (D): Proliferation of the control group and LAMP2A-overexpress group following 150 μM H2O2 treatment. (E): LAMP2A knockdown cell lines were exposed to H 2 O 2 (0, 150μM and 300μM), and the apoptosis rate was measured by FACS after 24-hour treatment. (F): LAMP2A over-expressing cells were exposed to H 2 O 2 (0, 150μM and 300μM), and the apoptosis rate was measured by FACS after 24-hour treatment. Note: L2A, LAMP2A; NC, negative control. Data are presented as mean±SD of three independent experiments. ns, no significance, *P < 0.05. **P < 0.01.

Article Snippet: The lentiviral vectors with shRNAs against LAMP2A were purchased from GeneChem Company, Shanghai, China (PIEL248064052).

Techniques: Control, Knockdown, Expressing, Negative Control

Journal: PLOS One

Article Title: LAMP2A-dependent chaperone-mediated autophagy enhances oxidative stress resistance in gastric cancer cells through selective degradation of accumulated oxidized DJ-1

doi: 10.1371/journal.pone.0331823

Figure Lengend Snippet: (A): Identification of a conserved KFERQ-like motif in DJ-1, a CMA substrate recognition signature. (B): WB analysis of DJ-1 protein expression levels. (C) H₂O₂ dose-dependent upregulation of DJ-1 protein in gastric cancer cells (150 μM, 300 μM; 24 h treatment). (D): Co-IP confirms enhanced DJ-1/LAMP2A interaction in H₂O₂-treated (300 μM, 24 h) MKN45 cancer cells. (E): IF demonstrates oxidative stress-induced colocalization of DJ-1 and LAMP2A in MKN45 cells (300 μM H₂O₂, 24 h). Data are presented as mean±SD of three independent experiments. ns, no significance, *P < 0.05. **P < 0.01.

Article Snippet: The lentiviral vectors with shRNAs against LAMP2A were purchased from GeneChem Company, Shanghai, China (PIEL248064052).

Techniques: Expressing, Co-Immunoprecipitation Assay

Journal: PLOS One

Article Title: LAMP2A-dependent chaperone-mediated autophagy enhances oxidative stress resistance in gastric cancer cells through selective degradation of accumulated oxidized DJ-1

doi: 10.1371/journal.pone.0331823

Figure Lengend Snippet: (A) MKN45shL2A and MKN45shNC were stimulated by H 2 O 2 (0,300 μm) for 24 hours, and the protein levels of LAMP2A, DJ-1, apoptosis-related proteins Bcl-2 and BAX were detected by WB. (B) The MKN45shL2A cells were transfected with the LAMP2A plasmid to generate the rescue group (sh L2A + LAMP2A). The protein levels of LAMP2A, DJ-1, Bcl-2, and BAX were analyzed by WB in all cell groups after a 24-hour exposure to 300 μM H₂O₂. Data are presented as mean±SD of three independent experiments. ns, no significance, *P < 0.05. **P < 0.01.

Article Snippet: The lentiviral vectors with shRNAs against LAMP2A were purchased from GeneChem Company, Shanghai, China (PIEL248064052).

Techniques: Transfection, Plasmid Preparation

Journal: PLOS One

Article Title: LAMP2A-dependent chaperone-mediated autophagy enhances oxidative stress resistance in gastric cancer cells through selective degradation of accumulated oxidized DJ-1

doi: 10.1371/journal.pone.0331823

Figure Lengend Snippet: ①CMA inhibits tumor cell apoptosis through selective removal of overoxidized DJ-1 protein. ②In pathological conditions, ROS-mediated overoxidation converts DJ-1 into a pro-apoptotic form that triggers gastric cancer cell death, distinct from its physiological oxidative modification under basal oxidative stress. ③Oxidative stress serves as the primary inducer of LAMP2A upregulation in gastric cancer cells, establishing a compensatory mechanism for CMA activation.

Article Snippet: The lentiviral vectors with shRNAs against LAMP2A were purchased from GeneChem Company, Shanghai, China (PIEL248064052).

Techniques: Modification, Activation Assay