rabbit anti lc3a b  (Proteintech)

Journal: Investigative ophthalmology & visual science

Article Title: PERK Regulates Epithelial-Mesenchymal Transition Through Autophagy and Lipid Metabolism in Lens Epithelial Cells.

doi: 10.1167/iovs.66.5.35

Figure Lengend Snippet: FIGURE 8. ISRIB attenuates TGFβ2-mediated autophagy. (A) Representative transmission electron microscopy (TEM) images of LECs exposed to TGFβ2 and ISRIB for 48 hours. Red arrows indicate autophagic vacuoles. (B) LECs were infected with the mRFP-GFP-LC3 reporter virus and exposed to TGFβ2 and ISRIB for 48 hours (red: autolysosomes; yellow: autophagosomes). Scale bar: 10 μm. (C) Immunofluorescence staining revealed P62 expression in LECs exposed to TGFβ2 and ISRIB for 48 hours. Scale bar: 50 μm. (D) Western blot analysis of P62 and α-SMA expression levels after cotreatment with TGFβ2 and ISRIB (4 μM) at different time points. (E) Western blot analysis of LC3 and P62 expression levels after cotreatment with TGFβ2 and different concentrations of ISRIB.

Article Snippet: The primary antibodies (1:1000) used in Western blot analysis were as follows: GAPDH (2118; CST), BiP (3177; CST), ACTIN (3700; CST), p-eIF2α (9721; CST), eIF2α (9722; CST), IRE1α (3294; CST), vimentin (5741; CST), βcatenin (8480; CST), LC3 (12741; CST), p-SMAD2/3 (8828; CST), SMAD2/3 (8685; CST), FN (ab137720; Abcam), α-SMA (ab7817; Abcam), and P62 (ab56416; Abcam).

Techniques: Transmission Assay, Electron Microscopy, Infection, Virus, Immunofluorescence, Staining, Expressing, Western Blot

Journal: Investigative ophthalmology & visual science

Article Title: PERK Regulates Epithelial-Mesenchymal Transition Through Autophagy and Lipid Metabolism in Lens Epithelial Cells.

doi: 10.1167/iovs.66.5.35

Figure Lengend Snippet: FIGURE 9. Autophagy mediates the profibrotic effects of ISRIB. (A) Immunostaining of BiP and P62 in mouse subcapsular plaques in the control and ISRIB-treated groups. (B, C) FN and α-SMA expression levels were measured by quantitative RT-PCR and Western blotting in LECs treated with TGFβ2, ISRIB, and rapamycin (250 nM) for 72 hours. (D) Western blot analysis of LC3, P62, and α-SMA expression levels in LECs treated with TGFβ2, ISRIB, rapamycin, and CQ (50 μM) for 72 hours.

Article Snippet: The primary antibodies (1:1000) used in Western blot analysis were as follows: GAPDH (2118; CST), BiP (3177; CST), ACTIN (3700; CST), p-eIF2α (9721; CST), eIF2α (9722; CST), IRE1α (3294; CST), vimentin (5741; CST), βcatenin (8480; CST), LC3 (12741; CST), p-SMAD2/3 (8828; CST), SMAD2/3 (8685; CST), FN (ab137720; Abcam), α-SMA (ab7817; Abcam), and P62 (ab56416; Abcam).

Techniques: Immunostaining, Control, Expressing, Quantitative RT-PCR, Western Blot

Journal: Journal of translational medicine

Article Title: CircPRDM5-mediated regulation of miR-433-3p and HDAC6 in Parkinson's disease: a novel neuroprotective axis.

doi: 10.1186/s12967-025-06602-3

Figure Lengend Snippet: Fig. 3 Downregulation of circPRDM5 alleviates MPP+-induced cellular damage, apoptosis, and autophagy. A CCK8 cell viability assay for SH-SY5Y cells treated with 0, 0.5, 1, or 2 mM MPP+ for 24 h. B CCK8 cell viability assay for SH-SY5Y cells treated with 1 mM MPP+ at 0 h, 12 h, 24 h, or 48 h. C, D qRT-PCR analysis of circPRDM5 expression in SH-SY5Y cells treated with 0, 0.5, 1, or 2 mM MPP+ or in SH-SY5Y cells treated with 1 mM MPP+ at 0 h, 12 h, 24 h, or 48 h. E qRT-PCR analysis of circPRDM5 knockdown efficiency in SH-SY5Y cells of the sh-NC group and sh- circPRDM5 group without any treatment. F CCK8 cell viability assay in SH-SY5Y cells left untreated or treated with 1 mM of MPP+ for 48 h of the control group, MPP+ group, MPP+ + sh-NC group and MPP+ + sh-circPRDM5 group. G LDH levels in SH-SY5Y cells of the indicated groups. H, I Flow cytometry analysis was performed to assess apoptosis rate in SH-SY5Y cells of the indicated groups. J Western blot assay for apoptosis-related proteins in SH-SY5Y cells of the indicated groups. K Western blot assay for autophagy-related proteins in SH-SY5Y cells of the indicated groups. L Immunofluorescence staining for LC3 protein in SH-SY5Y cells of the indicated groups (scale bar = 50 μm). Data are shown as the mean ± SD of three independent experiments. P values were calculated using either unpaired Student’s t-test or one-way ANOVA followed by Bonferroni post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001, ##p < 0.01

Article Snippet: After protein transfer, the membranes were blocked with 5% non-fat dry milk in TBST (Tris-buffered saline with 0.1% Tween-20) and then incubated with the following primary antibodies overnight at 4 °C: Bax (5023S, 1:1000, Cell Signaling Technology, USA), Bcl-2 (2876S, 1:1000, Cell Signaling Technology, USA), LC3 I/II (12741S, 1:1000, Cell Signaling Technology, USA), p62/SQSTM1 (5114S, 1:1000, Cell Signaling Technology, USA), HDAC6 (7558S, 1:1000, Cell Signaling Technology, USA), and β-actin (3700, 1:1000, Cell Signaling Technology, USA) as a loading control.

Techniques: Viability Assay, Quantitative RT-PCR, Expressing, Knockdown, Control, Flow Cytometry, Western Blot, Immunofluorescence, Staining