Journal: BMC complementary medicine and therapies

Article Title: Piperine, a black pepper compound, induces autophagy and cellular senescence mediated by NF-κB and IL-6 in acute leukemia.

doi: 10.1186/s12906-024-04641-9

Figure Lengend Snippet: Fig. 2 NF-κB1 and IL-6 are piperine-responsive target proteins involved in autophagy and senescence. (A) All possible piperine-responsive target pro teins identified by the STITCH database. (B) Protein‒protein interactions of piperine-responsive target proteins and autophagy/senescence proteins, including mTOR, ULK1, CDKN1A (p21) and CDK2. The green lines represent proteins that directly interact with piperine. The red box indicates a group of proteins that interact with mTOR, ULK1, CDKN1A (p21) or CDK2. The blue box represents a group of autophagy and senescence proteins

Article Snippet: Primary antibodies against mTOR, CDK2, ULK1, p21 (Cell Signaling Technology, Danvers, MA, USA), Beclin-1, α-tubulin (Proteintech, USA) and NF-κB1 (Invitrogen, USA) were incubated with the PVDF membranes overnight at 4 °C.

Techniques:

Journal: BMC complementary medicine and therapies

Article Title: Piperine, a black pepper compound, induces autophagy and cellular senescence mediated by NF-κB and IL-6 in acute leukemia.

doi: 10.1186/s12906-024-04641-9

Figure Lengend Snippet: Fig. 3 Induction of autophagy by piperine is mediated by NF-κB1 in NB4 and MOLT-4 cells. NB4 and MOLT-4 cells were treated with piperine at the IC50 for 48 h. LC3 expression in piperine-treated NB4 and MOLT-4 cells was analyzed using flow cytometry. mTOR, ULK1 and NF-κB1 expression was determined using RT‒qPCR and western blotting. (A) Histogram of LC3-conjugated FITC fluorescence and the mean fluorescence intensity (MFI) of LC3 in piperine- treated NB4 and MOLT-4 cells. (B) mTOR, ULK1, BECN1 and NF-κB1 gene expression in piperine-treated NB4 and MOLT-4 cells. (C) Western blot showing the protein levels of mTOR, ULK1, Beclin-1 and NF-κB1 and the levels of the mTOR, ULK1, Beclin-1 and NF-κB1 proteins in piperine-treated NB4 and MOLT-4 cells. The data are expressed as the mean ± S.E.M. of three independent experiments. *p < 0.05, **p < 0.01 and ***p < 0.001 indicate statistically significant differences compared withthe control group

Article Snippet: Primary antibodies against mTOR, CDK2, ULK1, p21 (Cell Signaling Technology, Danvers, MA, USA), Beclin-1, α-tubulin (Proteintech, USA) and NF-κB1 (Invitrogen, USA) were incubated with the PVDF membranes overnight at 4 °C.

Techniques: Expressing, Flow Cytometry, Western Blot, Fluorescence, Gene Expression, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Silencing TUFM Inhibits Development of Monocrotaline-Induced Pulmonary Hypertension by Regulating Mitochondrial Autophagy via AMPK/mTOR Signal Pathway

doi: 10.1155/2022/4931611

Figure Lengend Snippet: Alleviated mitophagy by TUFM silence is associated with AMPK/mTOR pathway. (a) Representative western blot bands for TUFM, AMPK, p-AMPK, mTOR, p-mTOR, BECN1, Atg13, Atg16L1, ULK1, and Atg12. (b–k) Densitometry data represent the intensity of each group. The data is presented as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The related primary antibody included TUFM (Abcam, ab173300, 1 : 10000 dilution), LC3 (CST, #3868, 1 : 1000 dilution), BECN1 (CST, #3495, 1 : 1000 dilution), P62 (CST, #39749, 1 : 1000 dilution), ATG13 (Proteintech, #18258-1-AP, 1 : 1000 dilution), α -SMA (BOSTER, BM0002, 1 : 1000 dilution), CD31 (Abcam, ab222783, 1 : 2000 dilution), Bax (Abcam, ab182734, 1 : 1000 dilution), Bcl2 (HUABIO, ET1702-53, 1 : 1000 dilution), AMPK (Bioss, bs-5551R, 1 : 1000 dilution), p-AMPK (Bioss, bs-2771R, 1 : 1000 dilution), mTOR (Bioss, bs-3494R, 1 : 1000 dilution), p-mTOR (Bioss, bs-3495R, 1 : 1000 dilution), ULK1 (BOSTER, A00584-1, 1 : 1000 dilution), Apaf-1 (HUABIO, ET1607-12, 1 : 1000 dilution), ATG12 (CST, #4180, 1 : 1000 dilution), ATG16L1 (CST, #8089, 1 : 1000 dilution), and β -actin (CST, #4970, 1 : 1000 dilution) which was used as an internal reference.

Techniques: Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Silencing TUFM Inhibits Development of Monocrotaline-Induced Pulmonary Hypertension by Regulating Mitochondrial Autophagy via AMPK/mTOR Signal Pathway

doi: 10.1155/2022/4931611

Figure Lengend Snippet: Schematic figure of the current study. Increased TUFM expression in hypoxia-stimulated pulmonary arterial hypertension cells causes an increasing mtDNA translation, leading to dysfunction of the mitochondrial respiratory chain. Mitochondrial dysfunction induces cellular stress and then activates AMPK. On the one hand, activated AMPK decreases the phosphorylation level of mTOR, inhibits the activity of mTOR, and then disassociates from ULK1. Thus, phosphorylation of specific sites of ULK1 and Atg13 is released. Meanwhile, the ULK1 complex is activated through autophosphorylation at thr180 and phosphorylates Atg13, FIP200, atg101, and other Atg proteins. The activated ULK1 complex then translocates to the isolation membrane of the endoplasmic reticulum, where autophagy is initiated. On the other hand, activated AMPK will directly stimulate ULK1 and BECN1, initiating autophagy.

Article Snippet: The related primary antibody included TUFM (Abcam, ab173300, 1 : 10000 dilution), LC3 (CST, #3868, 1 : 1000 dilution), BECN1 (CST, #3495, 1 : 1000 dilution), P62 (CST, #39749, 1 : 1000 dilution), ATG13 (Proteintech, #18258-1-AP, 1 : 1000 dilution), α -SMA (BOSTER, BM0002, 1 : 1000 dilution), CD31 (Abcam, ab222783, 1 : 2000 dilution), Bax (Abcam, ab182734, 1 : 1000 dilution), Bcl2 (HUABIO, ET1702-53, 1 : 1000 dilution), AMPK (Bioss, bs-5551R, 1 : 1000 dilution), p-AMPK (Bioss, bs-2771R, 1 : 1000 dilution), mTOR (Bioss, bs-3494R, 1 : 1000 dilution), p-mTOR (Bioss, bs-3495R, 1 : 1000 dilution), ULK1 (BOSTER, A00584-1, 1 : 1000 dilution), Apaf-1 (HUABIO, ET1607-12, 1 : 1000 dilution), ATG12 (CST, #4180, 1 : 1000 dilution), ATG16L1 (CST, #8089, 1 : 1000 dilution), and β -actin (CST, #4970, 1 : 1000 dilution) which was used as an internal reference.

Techniques: Expressing, Phospho-proteomics, Activity Assay, Isolation, Membrane