Journal: Frontiers in pharmacology

Article Title: Shengma Biejia Decoction Inhibits Cell Growth in Multiple Myeloma by Inducing Autophagy-Mediated Apoptosis Through the ERK/mTOR Pathway.

doi: 10.3389/fphar.2021.585286

Figure Lengend Snippet: FIGURE 3 | SMBJD is safe and could inhibit the expression of autophagy-related proteins, promote apoptosis-related proteins in vivo. (A) Representative pictures of peer group mouse liver, spleen, and kidney after HE staining (× 200), scale bar 100 µm. (B) The expression levels of P62, LC3, and cleaved caspase-3 in transplanted tumors were detected using immunohistochemistry (× 200), scale bar 100 µm. (C) Extracts from transplanted tumors were analyzed for autophagy- related and apoptosis-related proteins expression by western blot. (D) The corresponding expression of P62, LC3, and cleaved caspase-3 levels in tumor tissue are shown as histograms. (E) The corresponding autophagy-related and apoptosis-related proteins expression by western blot are shown as histograms. (*p < 0.05 compared with the model group).

Article Snippet: The membranes were blocked with 5% fat-free milk with 0.1% Tween-20 in 0.02 ml of TBS buffer (TBST) for 1 h and incubated with primary antibodies against P62 (1:1,000–4,000, Proteintech, IL, United States), LC3 (1:600–2,500, Proteintech, IL, United States), Bax (1:1,000, CST, MA, United States), Bcl-2 (1:1,000, CST, MA, United States), caspase-3 (1:1,000, CST, MA, United States), cleaved caspase-3 (1:1,000, CST,MA, United States), ERK1/2 (1:10,000, Abcam, Cambridge, United Kingdom), phosphorylated ERK1/2 (1:5,000–10,000, Abcam, Cambridge, United Kingdom), Ser2448 mTOR (1:500–2,000, Sciben, Nanjing, China), and phosphorylated Ser2448 mTOR (1: 500–2,000, Sciben, Nanjing, China) at 4°C overnight.

Techniques: Expressing, In Vivo, Staining, Immunohistochemistry, Western Blot

Journal: Frontiers in pharmacology

Article Title: Shengma Biejia Decoction Inhibits Cell Growth in Multiple Myeloma by Inducing Autophagy-Mediated Apoptosis Through the ERK/mTOR Pathway.

doi: 10.3389/fphar.2021.585286

Figure Lengend Snippet: FIGURE 4 | SMBJD represses the growth, inhibits autophagy, and promotes apoptosis of H929 and U266 cells. (A) The cell viability of H929 and U266 cells after different concentrations and time of SMBJD treatment were analyzed with MTT assay. (B) IC50 value of H929 and U266 cells after SMBJD intervention. (C) H929 and U266 cells were transiently transfected with Ad-GFP-LC3 for 24 h. Then the cells were cultured with SMBJD for another 36 h. The formation of GFP-LC3 puncta were examined by a confocal microscope and typical images were presented (× 400), scale bar 2 µm. (D) Annexin V/PI staining of H929 and U266 cells apoptosis rate treated with SMBJD for 36 h (E, F) The quality graphs of C and D. Data are expressed as the means ± SD (nsP > 0.05 compared with each other, *p < 0.05 compared with control group).

Article Snippet: The membranes were blocked with 5% fat-free milk with 0.1% Tween-20 in 0.02 ml of TBS buffer (TBST) for 1 h and incubated with primary antibodies against P62 (1:1,000–4,000, Proteintech, IL, United States), LC3 (1:600–2,500, Proteintech, IL, United States), Bax (1:1,000, CST, MA, United States), Bcl-2 (1:1,000, CST, MA, United States), caspase-3 (1:1,000, CST, MA, United States), cleaved caspase-3 (1:1,000, CST,MA, United States), ERK1/2 (1:10,000, Abcam, Cambridge, United Kingdom), phosphorylated ERK1/2 (1:5,000–10,000, Abcam, Cambridge, United Kingdom), Ser2448 mTOR (1:500–2,000, Sciben, Nanjing, China), and phosphorylated Ser2448 mTOR (1: 500–2,000, Sciben, Nanjing, China) at 4°C overnight.

Techniques: MTT Assay, Transfection, Cell Culture, Microscopy, Staining, Control

Journal: Frontiers in pharmacology

Article Title: Shengma Biejia Decoction Inhibits Cell Growth in Multiple Myeloma by Inducing Autophagy-Mediated Apoptosis Through the ERK/mTOR Pathway.

doi: 10.3389/fphar.2021.585286

Figure Lengend Snippet: FIGURE 5 | SMBJD inhibits autophagy-related proteins and promotes apoptosis-related proteins of multiple myeloma cell lines H929 and U266 (A) Western blot analysis of autophagy-related and apoptosis-related proteins expression after SMBJD administration for 36 h in both cells. (B) Western blot analysis of LC3 expression after SMBJD, 3-MA, Baf-A1, and SMBJD + Baf-A1 administration for 36 h in H929 and U266 cells.

Article Snippet: The membranes were blocked with 5% fat-free milk with 0.1% Tween-20 in 0.02 ml of TBS buffer (TBST) for 1 h and incubated with primary antibodies against P62 (1:1,000–4,000, Proteintech, IL, United States), LC3 (1:600–2,500, Proteintech, IL, United States), Bax (1:1,000, CST, MA, United States), Bcl-2 (1:1,000, CST, MA, United States), caspase-3 (1:1,000, CST, MA, United States), cleaved caspase-3 (1:1,000, CST,MA, United States), ERK1/2 (1:10,000, Abcam, Cambridge, United Kingdom), phosphorylated ERK1/2 (1:5,000–10,000, Abcam, Cambridge, United Kingdom), Ser2448 mTOR (1:500–2,000, Sciben, Nanjing, China), and phosphorylated Ser2448 mTOR (1: 500–2,000, Sciben, Nanjing, China) at 4°C overnight.

Techniques: Western Blot, Expressing

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: Exosomes derived from fibroblasts in DFUs delay wound healing by delivering miR-93-5p to target macrophage ATG16L1.

doi: 10.1016/j.bbadis.2024.167640

Figure Lengend Snippet: Fig. 3. miR-93-5p affects macrophage autophagy function by affecting Atg16L1. (A) Nine mRNAs were identifed based based on TargetScan, miRDB, miRTarBase and Human Autophage Database. (B) Schematic representation of the wild-type and mutant-type binding site between the 3′UTR of ATG16L1 and miR-93-5p. (C) Relative luciferase activity of 3′UTR-ATG16L1-luc constructs in HEK293T cells after transfection of miR-93-5p mimics/NC; n = 3 per group. (D) The protein levels of ATG16L1 in bone marrow-derived macrophages transfected with miR-93-5p NC/mimics or PBS (Control) by Western blot. (E) The protein levels of ATG16L1 in skin tissues from normal or diabetic wound (DFUs) by Western blot. (F) Immunofluorescence staining for ATG16L1 (red) and F4/80 (green) in skin tissues from normal or diabetic wound (DFUs); scale bar, 100 μm. (G) The protein levels of LC3b and P62 in BMDMs transfected with miR-93-5p NC/mimics or PBS by Western blot, immunofluorescence staining for LC3b (green) in bone marrow-derived macrophages transfected with miR-93-5p NC/mimics or PBS; scale bar, 50 μm. (H) The protein levels of ATG16L1, LC3 and P62 in fibroblast-derived exosomes treated bone marrow-derived macrophages transfected with miR-93-5p inhibitor NC/in hibitor or PBS (Control) by Western blot. Data are expressed as mean ± SD, ns P > 0.05, **P < 0.01 (unpaired t-test or ANOVA). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Next, sections were incubated overnight at 4 ◦C with primary antibodies against F4/80 (Servicebio, GB11027), Atg16L1 (CST, #8089), LC3B (Proteintech, 81004-1-RR), α-SMA (Proteintech,14395-1AP) and Collagen Type I (Proteintech, 14695-1-AP).

Techniques: Mutagenesis, Binding Assay, Luciferase, Activity Assay, Construct, Transfection, Derivative Assay, Control, Western Blot, Immunofluorescence, Staining

Journal: Autophagy

Article Title: BIRC5/Survivin is a novel ATG12–ATG5 conjugate interactor and an autophagy-induced DNA damage suppressor in human cancer and mouse embryonic fibroblast cells

doi: 10.1080/15548627.2019.1671643

Figure Lengend Snippet: BIRC5 downregulation induces autophagy-dependent DNA damage in human cancer and mouse embryonic fibroblast cells. (A) MCF7 and MDA-MB-231 cells were treated with or without 2xIC 50 YM155 and co-incubated with or without CQ for 48 h. DNA damage was detected using comet assay. A statistically significant difference ( P < 0.0001) in the relative tail moment of cells treated with YM155 versus YM155 + CQ is denoted by a “***”. (B and C) MCF7, MDA-MB-231, and MEF cells were transfected with either scramble siRNA or BIRC5 ( Birc5 ) siRNA and co-incubated with or without CQ (15 μM), 3MA (4 mM), and BAF (3 nM) for 48 h. DNA damage was detected using comet assay. A statistically significant difference ( P < 0.0001) in the relative tail moment of cells treated with BIRC5 ( Birc5 ) siRNA versus BIRC5 (Birc5) siRNA + CQ/3MA/BAF is denoted by a “***”. (D) MDA-MB-231 and MEF cells were transfected with either scramble siRNA or BIRC5 ( Birc5 ) siRNA and co-incubated with or without CQ (15 μM) or LC3B ( Lc3b ) siRNA transfection for 48 h. Expression of p-H2AX was examined by western blot analysis. (E) MEF and atg5 −/- MEF cells were transfected with either scramble siRNA or Birc5 siRNA for 48 h. Expression of p-H2AX was examined by western blot analysis. Scale bars: 50 μm (A, B, and C).

Article Snippet: The resolved proteins were transferred onto a PVDF membrane (Millipore, IPVH00010), which was then exposed to 5% nonfat dried milk (Fonterra)/bovine serum albumin (BSA; Sigma-Aldrich, A2153) in TBST buffer (2.44 g/L Tris base [Calbiochem, 9210], 8.76 g/L NaCl [Calbiochem, 567441], 0.05% Tween® 20 [Calbiochem, 9480-OP], pH 7.4) for 1 h at room temperature before incubation overnight at 4°C with primary antibodies: anti-BIRC5 (R&D Systems, AF886); anti-ATG7 (Millipore, AB10511); anti-ATG12 (Gene Tex, GTX124181); anti-ATG5 (Millipore, MAB2605); anti-ATG16L1 (Millipore, ABC25); anti-LC3B (Origene, TA301543); anti-SQSTM1 (Gene Tex, GTX100685); anti-p-MTOR (Ser2448) (Cell Signaling, 2971); anti-p-H3-3A (Abcam, ab32107); anti-ACTA1/actin (Millipore, MAB1501).

Techniques: Incubation, Single Cell Gel Electrophoresis, Transfection, Expressing, Western Blot

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: Frontiers in Molecular Biosciences

Article Title: Exploring the interaction mechanisms of CD46/TREM1 and LC3B/ATG5 in the inflammation-cancer transformation of oral squamous cell carcinoma based on bioinformatics

doi: 10.3389/fmolb.2025.1713632

Figure Lengend Snippet: Expression of CD46/TREM1 and LC3B/ATG5 in four stages of SD rats: normal (NC), inflammatory (INF), leukoplakia (OLK), and squamous cell carcinoma (OSCC) (×20).

Article Snippet: Primary antibodies used: CD46 (1:500, MCE, HY-P 80064), TREM1 (1:300, Bioss, bs-10306R), LC3B (1:100, MCE, HY-P 80742), ATG5 (1:600, Immunoway, YM8340).

Techniques: Expressing

Journal: Frontiers in Molecular Biosciences

Article Title: Exploring the interaction mechanisms of CD46/TREM1 and LC3B/ATG5 in the inflammation-cancer transformation of oral squamous cell carcinoma based on bioinformatics

doi: 10.3389/fmolb.2025.1713632

Figure Lengend Snippet: Expression of CD46/TREM1 and LC3B/ATG5 in human normal (NC), leukoplakia (OLK), and squamous cell carcinoma (OSCC) tissues (×20).

Article Snippet: Primary antibodies used: CD46 (1:500, MCE, HY-P 80064), TREM1 (1:300, Bioss, bs-10306R), LC3B (1:100, MCE, HY-P 80742), ATG5 (1:600, Immunoway, YM8340).

Techniques: Expressing