rabbit polyclonal antibody against lamp2a  (Abcam)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Identification of a novel aromatic-turmerone analog that activates chaperone-mediated autophagy through the persistent activation of p38

doi: 10.3389/fcell.2024.1418296

Figure Lengend Snippet: Primer sequences used in RT-qPCR analyses.

Article Snippet: The anti-human LAMP2A rabbit polyclonal antibody (ab125068), anti-TSG101 rabbit polyclonal antibody (ab125011), and anti-α-synuclein rabbit monoclonal antibody (ab138501) were obtained from AbCam (Cambridge, United Kingdom).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Identification of a novel aromatic-turmerone analog that activates chaperone-mediated autophagy through the persistent activation of p38

doi: 10.3389/fcell.2024.1418296

Figure Lengend Snippet: Effects of ar-turmerone analogs on CMA/mA activity in SH-SY5Y cells stably expressing GAPDH-HT. (A) Representative fluorescence image of SaraFluor 650T attached to GAPDH-HT in cells treated with vehicle, MPA (5 μM), or ar-turmerone analogs (20 μM) for 24 h. (C) Representative fluorescence image of GFP and SaraFluor 650T attached to GAPDH-HT and merged images of GFP (green) and SaraFluor (magenta) in cells treated with vehicle or A4 (20 μM) for 24 h. Cells expressing GFP means cells expressing miRNA (negative control miRNA, LAMP2A miRNA, or TSG101 miRNA). Scale bars = 10 μm. (B, D) Quantitative analyses of GAPDH-HT puncta per cell shown in A and C, respectively. Data are represented as the percentage of GAPDH-HT puncta in cells treated with vehicle (B) and in cells transfected with control miRNA and treated with vehicle (D) . Numbers in the columns represent the number of observed cells. n.s., not significant, *** p < 0.001 (Kruskal–Wallis test, followed by a post hoc Dunn’s test).

Article Snippet: The anti-human LAMP2A rabbit polyclonal antibody (ab125068), anti-TSG101 rabbit polyclonal antibody (ab125011), and anti-α-synuclein rabbit monoclonal antibody (ab138501) were obtained from AbCam (Cambridge, United Kingdom).

Techniques: Activity Assay, Stable Transfection, Expressing, Fluorescence, Negative Control, Transfection, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Identification of a novel aromatic-turmerone analog that activates chaperone-mediated autophagy through the persistent activation of p38

doi: 10.3389/fcell.2024.1418296

Figure Lengend Snippet: Effects of ar-turmerone analogs on the expression of CMA-related proteins and mRNAs in SH-SY5Y cells stably expressing GAPDH-HT. (A) Representative immunoblot images of LAMP2A, Hsc70, MEF2D, and β-tubulin in cells treated with vehicle, 5 μM MPA, or ar-turmerone analogs (20 μM) for 24 h. (B) Quantitative analyses of the immunoreactive bands of LAMP2A, Hsc70, and MEF2D. The band intensities were normalized with the bands of β-tubulin as an internal control. Data are presented as the mean ± SEM of four different samples. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. vehicle (one-way ANOVA, followed by a post hoc Tukey test). (C) The mRNA amounts of LAMP2A, LAMP2B, LAMP1, HSPA8 (Hsc70), TFEB, and MEF2D were quantified with RT-qPCR. Cells were treated with vehicle, 20 μM A2, or 20 μM A4 for 24 h. The data are presented as the mean ± SEM of four independent samples. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. vehicle (one-way ANOVA, followed by a post hoc Tukey test).

Article Snippet: The anti-human LAMP2A rabbit polyclonal antibody (ab125068), anti-TSG101 rabbit polyclonal antibody (ab125011), and anti-α-synuclein rabbit monoclonal antibody (ab138501) were obtained from AbCam (Cambridge, United Kingdom).

Techniques: Expressing, Stable Transfection, Western Blot, Control, Quantitative RT-PCR

Journal: Frontiers in Cell and Developmental Biology

Article Title: Identification of a novel aromatic-turmerone analog that activates chaperone-mediated autophagy through the persistent activation of p38

doi: 10.3389/fcell.2024.1418296

Figure Lengend Snippet: Effects of ar-turmerone analogs on the cytotoxicity of rotenone in SH-SY5Y cells stably expressing GAPDH-HT. (A) Survival of cells treated with vehicle and rotenone (10 μM) and co-treated with rotenone and A2 (2, 20 μM) or A4 (2, 20 μM). (B) Survival of cells treated with vehicle, A4 (20 μM), and/or SB203580 (SB, 10 μM) in the absence or presence of rotenone (10 μM). Data are represented as the percentage of cell survival in cells treated with vehicle alone and as the mean ± SEM of twelve (A) and fourteen (B) different samples. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. vehicle alone, # p < 0.05, ## p < 0.01 (Kruskal–Wallis test, followed by a post hoc Dunn’s test). (C) Representative fluorescence image of GFP and DAPI and merged images of GFP (green) and DAPI (blue) in cells transfected with plasmids to express GFP and control or LAMP2A miRNA. Cells were treated with vehicle (Veh, 0.2% DMSO), 10 μM rotenone (Rot), or rotenone and 20 μM A4 (Rot + A4) for 24 h. Yellow arrows indicate the GFP-positive cells with condensed nuclei. Scale bars = 50 μm. (D) Quantitative analyses of cytotoxicity indicated by the nuclear condensation. We calculated the percentage of cells with condensed nuclei in all GFP-positive cells. Data are presented as the mean ± SEM of four different wells. n.s.: not significant, * p < 0.05, *** p < 0.001 (one way ANOVA, followed by a post hoc Tukey test).

Article Snippet: The anti-human LAMP2A rabbit polyclonal antibody (ab125068), anti-TSG101 rabbit polyclonal antibody (ab125011), and anti-α-synuclein rabbit monoclonal antibody (ab138501) were obtained from AbCam (Cambridge, United Kingdom).

Techniques: Stable Transfection, Expressing, Fluorescence, Transfection, Control