Journal: Endocrinology

Article Title: Deficiency of C1QL1 reduced murine ovarian follicle reserve through intraovarian and endocrine control.

doi: 10.1210/endocr/bqac048

Figure Lengend Snippet: Figure 5. Deficiency of C1QL1 promoted apoptosis and inhibited autophagy in the ovary. (A) Representative images of TUNEL staining of ovarian sections. The green signals indicate representative cells with positive TUNEL staining, which was mainly detected in the large atretic follicles. (B) The intensity of TUNEL-positive signals was shown in charts (n = 4 per group). (C) Representative images of immunofluorescence staining of ovarian sections with LC3B antibody. LC3B-positive signals were found in granulosa cells and oocytes and were weakened in C1QL1-deficient ovaries. (D) The intensity of LC3B-positive signals are shown (n = 4 per group). (E) Apoptosis- and autophagy-related proteins in the ovaries were analyzed by Western blotting. Representative blots are shown in the left panel, and the amount of protein normalized to β-tubulin, which is presented as mean ± SEM, is shown in the right panel. n = 6 per group. *P < 0.05, **P < 0.01, ***P < 0.001. NS, no significance.

Article Snippet: The sections and granulosa cells were stained with anti-rabbit LC3B antibody and Alexa Fluor 594-conjugated secondary antibody and mounted with DAPI (Boster).

Techniques: TUNEL Assay, Staining, Immunofluorescence, Western Blot

Journal: Human & experimental toxicology

Article Title: Hypoxia-induced autophagy attenuates ferredoxin 1-mediated cuproptosis in colorectal cancer cells.

doi: 10.1177/09603271251335393

Figure Lengend Snippet: Figure 4. Hypoxia induces autophagy in colorectal cancer cells. (a) Western blot analysis of LC3B, Beclin 1, and P62 expression in LoVo and RKO cells under hypoxic conditions. (b) Immunofluorescence staining of LC3B in LoVo and RKO cells under hypoxic conditions. *p < .05, **p < .01.

Article Snippet: Tissue sections were deparaffinized and incubated with the LC3B antibody (BM4827, 1:200; Boster, Shanghai, China) and secondary antibodies.

Techniques: Western Blot, Expressing, Staining

Journal: Human & experimental toxicology

Article Title: Hypoxia-induced autophagy attenuates ferredoxin 1-mediated cuproptosis in colorectal cancer cells.

doi: 10.1177/09603271251335393

Figure Lengend Snippet: Figure 5. Hypoxia-induced autophagy attenuates FDX1-mediated cuproptosis in colorectal cancer cells. (a) Western blot analysis of LC3B, Beclin 1, and P62 expression in LoVo and RKO cells treated with rapamycin under hypoxic conditions. (b) Immunofluorescence staining of LC3B in LoVo and RKO cells. (c) Transwell assay of cell migration and invasion in LoVo and RKO cells. (d) EdU staining and TUNEL assay of cell proliferation and damage in LoVo and RKO cells. (e) Western blot analysis of DLAT oligomerization in LoVo and RKO cells. *p < .05, **p < .01.

Article Snippet: Tissue sections were deparaffinized and incubated with the LC3B antibody (BM4827, 1:200; Boster, Shanghai, China) and secondary antibodies.

Techniques: Western Blot, Expressing, Staining, Transwell Assay, Migration, TUNEL Assay

Journal: Human & experimental toxicology

Article Title: Hypoxia-induced autophagy attenuates ferredoxin 1-mediated cuproptosis in colorectal cancer cells.

doi: 10.1177/09603271251335393

Figure Lengend Snippet: Figure 6. Autophagy inhibition enhances the tumor-suppressive effects of FDX1 overexpression in vivo. (a) Representative images of tumors from nude mice implanted with LoVo cells under different conditions. (b, c, d) Tumor volume (b, c) and weight of nude mice implanted with LoVo cells under different conditions. (e) H&E staining and immunohistochemical staining of Ki67 in tumor tissues from nude mice implanted with LoVo cells under different conditions. (f) TUNEL assay of cell damage in tumor tissues from nude mice implanted with LoVo cells under different conditions. (g) qPCR analysis of FDX1 expression in tumor tissues from nude mice implanted with LoVo cells under different conditions. (g) Western blot analysis of FDX1 and DLAT oligomerization in tumor tissues. (h) Western blotting of LC3B and Beclin 1 expression in tumor tissues from nude mice implanted with LoVo cells under different conditions. *p < .05, **p < .01, ns noted no significance.

Article Snippet: Tissue sections were deparaffinized and incubated with the LC3B antibody (BM4827, 1:200; Boster, Shanghai, China) and secondary antibodies.

Techniques: Inhibition, Over Expression, In Vivo, Staining, Immunohistochemical staining, TUNEL Assay, Expressing, Western Blot

Journal: Cell Death & Disease

Article Title: Proton pump inhibition induces autophagy as a survival mechanism following oxidative stress in human melanoma cells

doi: 10.1038/cddis.2010.67

Figure Lengend Snippet: Effects of ESOM on autophagic flux. ( a ) Me30966 cells treated with ESOM (160 μ M) for 8 h are characterized by massive intracellular vacuolization. ( b ) Treatment with ESOM (for 8 h) induces accumulation of LC3-positive autophagic vacuoles in Me30966 and WM793 cells. ( c ) ESOM induces accumulation of the LC3-II form in autophagy-competent melanoma cells, as shown by the further accumulation of LC3-II in cells pretreated with inhibitors of lysosomal proteases. ( d ) ESOM-induced alteration of the autophagic flux is confirmed by accumulation of p62. ( e and f ) Analysis of the mRFP-GFP-LC3-plasmid shows the accumulation of autophagosomes in ESOM-treated Me30966 cells. Cells cultured in EBSS were used as positive control

Article Snippet: Anti-LC3 antibody against a synthetic LC3 peptide was produced in rabbits by NeoMPS (Strasbourg, France).

Techniques: Plasmid Preparation, Cell Culture, Positive Control

Journal: Cell Death & Disease

Article Title: Proton pump inhibition induces autophagy as a survival mechanism following oxidative stress in human melanoma cells

doi: 10.1038/cddis.2010.67

Figure Lengend Snippet: Role of ROS and mTOR in ESOM effects on the autophagic flux. ( a and b ) NAC pretreatment prevents the accumulation of autophagosomes induced by ESOM (160 μ M) in Me30966 cells, as shown by confocal fluorescence analysis of endogenous LC3 and quantification of cells with LC3+ dots. ( c ) Western blot analysis of LC3-I and LC3-II in the presence or absence of NAC in ESOM-treated cells. ( d and e ) ESOM treatment inhibits the mTOR pathway as shown by the time-dependent reduced phosphorylation of 4-EBP1 and p70S6K proteins. Such inhibition is prevented by the ROS scavenger NAC

Article Snippet: Anti-LC3 antibody against a synthetic LC3 peptide was produced in rabbits by NeoMPS (Strasbourg, France).

Techniques: Fluorescence, Western Blot, Phospho-proteomics, Inhibition

Journal: Cell Death & Disease

Article Title: Proton pump inhibition induces autophagy as a survival mechanism following oxidative stress in human melanoma cells

doi: 10.1038/cddis.2010.67

Figure Lengend Snippet: Autophagy is a cytoprotective mechanism in ESOM-treated melanoma cells. ( a ) Me30966 cells pretreated with Baf-A1 (40 nM) show increased sensitivity to ESOM-induced apoptosis in association with a further increased LC3-II accumulation. ( b ) Knockdown of Atg5 and beclin-1 induces a cytosolic localization of LC3 as compared with autophagosomal localization in control Me30966 cells. ( c and d ) Knockdown of Atg5 and beclin-1 increases the cytotoxicity of ESOM in Me30966 cells and WM793 cells, respectively

Article Snippet: Anti-LC3 antibody against a synthetic LC3 peptide was produced in rabbits by NeoMPS (Strasbourg, France).

Techniques: Knockdown, Control

Journal: Cells

Article Title: Enhanced Autophagic Flux, Suppressed Apoptosis and Reduced Macrophage Infiltration by Dasatinib in Kidneys of Obese Mice

doi: 10.3390/cells11040746

Figure Lengend Snippet: Sample size calculation.

Article Snippet: After blocking, an overnight membrane incubation with the primary antibodies—rabbit polyclonal antibody for LC3 and rabbit polyclonal antibody for SQSTM1/P62 as markers for autophagic activity, mouse monoclonal antibody for caspase-3 as an apoptosis marker and β-actin as a control protein (Biospes; YPA1652, ABclonal; A11250, Santa Cruz sc-7272 and Abcam ab227387, respectively)—was performed overnight at 4 °C (1:500, 1:1000, 1:100 and 1:5000 dilutions, respectively).

Techniques:

Journal: Cells

Article Title: Enhanced Autophagic Flux, Suppressed Apoptosis and Reduced Macrophage Infiltration by Dasatinib in Kidneys of Obese Mice

doi: 10.3390/cells11040746

Figure Lengend Snippet: ( A – D ) Protein analysis by Western blotting. ( A ) LC3 I and II (Molecular weight: 18 kDa and 16 kDa, respectively). ( B ) p62 (Molecular weight: 62 kDa). ( C ) Caspase-3: 32 kDa). ( D ) β-actin: 42 kDa). ( E – H ): LC3II, LC3II/LC3I ratio, P62 and caspase3 proteins relative quantitation /β-actin ratio) by Western blotting. Data are mentioned as mean ± SE; different letters = significant difference). LC3: Microtubule-associated proteins 1A/1B light chain 3B. M: Marker. MW: molecular weight.

Article Snippet: After blocking, an overnight membrane incubation with the primary antibodies—rabbit polyclonal antibody for LC3 and rabbit polyclonal antibody for SQSTM1/P62 as markers for autophagic activity, mouse monoclonal antibody for caspase-3 as an apoptosis marker and β-actin as a control protein (Biospes; YPA1652, ABclonal; A11250, Santa Cruz sc-7272 and Abcam ab227387, respectively)—was performed overnight at 4 °C (1:500, 1:1000, 1:100 and 1:5000 dilutions, respectively).

Techniques: Western Blot, Molecular Weight, Quantitation Assay, Marker

Journal: Cells

Article Title: Enhanced Autophagic Flux, Suppressed Apoptosis and Reduced Macrophage Infiltration by Dasatinib in Kidneys of Obese Mice

doi: 10.3390/cells11040746

Figure Lengend Snippet: ( A – D ): Immunohistochemical staining for LC3 (×400) in the kidney tissues of vehicle, Dasatinib control, Obesogenic diet and Obesogenic diet + Dasatinib groups ( A – D , respectively). Scale bar = 50 µm. ( E ) The mean percentage of LC3 immunopositive area. Results are presented as mean ± standard error. Different letters mean significant difference. Arrows: LC3 + ve cells. LC3 = Microtubule-associated protein light chain 3.

Article Snippet: After blocking, an overnight membrane incubation with the primary antibodies—rabbit polyclonal antibody for LC3 and rabbit polyclonal antibody for SQSTM1/P62 as markers for autophagic activity, mouse monoclonal antibody for caspase-3 as an apoptosis marker and β-actin as a control protein (Biospes; YPA1652, ABclonal; A11250, Santa Cruz sc-7272 and Abcam ab227387, respectively)—was performed overnight at 4 °C (1:500, 1:1000, 1:100 and 1:5000 dilutions, respectively).

Techniques: Immunohistochemical staining, Staining, Control