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

Journal: Physiological Reports

Article Title: Moderate alcohol consumption does not impair overload-induced muscle hypertrophy and protein synthesis

doi: 10.14814/phy2.12333

Figure Lengend Snippet: Markers of autophagy were differentially regulated by alcohol in response to muscle overload. The relative amount of phosphorylated and total ULK1 (A, B), total p62 (C), and LC3-II (D) were determined in the plantaris muscle 14-days after muscle overload. Representative images of each marker are provided along with a representative image for GAPDH which was used for normalization of p62 and LC3A/B-II expression. * P < 0.05, indicates difference from Sham condition within that treatment; ^ P < 0.05, indicates difference from Control-Sham. Values are expressed as means ± SE.

Article Snippet: Antibodies included (Cell Signaling, Beverly, MA, unless otherwise noted): S6K1, S6K1 (Thr 389 ), rpS6, rpS6 (Ser 240/244 ), 4E-BP1 (Bethyl Laboratories, Montgomery, TX), 4E-BP1 (Thr 37/46 ), eEF2, eEF2 (Thr 56 ), ERK1/2 (Thr 202 /Tyr 204 ), p42/44 MAPK, mTOR, mTOR (Ser 2448 ), REDD1 (ProteinTech, Chicago, IL), Akt, Akt (Thr 308 ), PRAS40, PRAS40 (Thr 246 ), ULK1, ULK1 (Ser 757 ), p62, LC3A/B, and GAPDH.

Techniques: Marker, Expressing