Journal: Biology Open

Article Title: Targeted inhibition of GRP78 by HA15 promotes apoptosis of lung cancer cells accompanied by ER stress and autophagy

doi: 10.1242/bio.053298

Figure Lengend Snippet: Inhibition of GRP78 by HA15 triggered autophagy. (A) The representative electron microscopic picture of A549 cell without HA15 treatment. (B) The representative autophagosome electron microscopic picture of A549 treated with 10 μM HA15 for 24 h. The autophagic vacuoles are indicated by red arrows. (C) The expressions of Atg5 , Atg7 , Atg12 , LC3 and ULK1 in A549 cells treated with 10 μM HA15 for 24 h tested by RT-PCR. (D) The expressions of Beclin1, p62, LC3B in A549 cells treated with 10 μM HA15 for 24 h tested by western blot. (E) The cell viability of A549 cells after treated with 10 μM chloroquine (CHQ) for 6 h and 10 μM HA15 for 24 h. n =3 individual experiments, *P <0.05, **P <0.01, ***P <0.001.

Article Snippet: To check the effect of autophagy on cell viability, autophagy inhibitor chloroquine (CHQ, MCE, USA) was used to treat A549 cells together with HA15.

Techniques: Inhibition, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Antioxidants

Article Title: Differential Effects of Three Medium-Chain Fatty Acids on Mitochondrial Quality Control and Skeletal Muscle Maturation

doi: 10.3390/antiox13070821

Figure Lengend Snippet: Effects of autophagy enhancement by C8 on skeletal muscle differentiation. C2C12 cells were induced to myotube differentiation for 48 h with pretreatment with C8 (48 h). We then examined the effects of C8 on skeletal muscle differentiation when autophagy was inhibited by chloroquine (CQ). ( A ) MyoD expression. ( B ) Semi-quantification of panel ( A ). ( C ) Fluorescence immunostaining for MyoD. ( D ) MyoD-positive rate. ( E ) MYH levels. ( F ) Semi-quantification of MYH protein content normalized to cell number. ( G ) MYH fluorescent fluorescence. ( H ) MYH-positive cell area. ( I ) Fused cell ratio. Scale bar, 50 μm; error bar, standard deviation from three independent trials. Statistical differences were calculated using analysis of variance (ANOVA) with Bonferroni correction. C8, caprylic acid; CQ, chloroquine; MyoD, myogenic differentiation 1; MYH, myosin heavy chain; CTRL, control; C8, caprylic acid; Hoechst, Hoechst 33342 dye.

Article Snippet: C2C12 myoblasts were pretreated with C8 (200 μg/mL) and chloroquine (CQ, 10 μM, HY-17589, Med Chem Express, Princeton, NJ, USA) for 48 h and then treated using the differentiation procedure described above.

Techniques: Expressing, Fluorescence, Immunostaining, Standard Deviation, Cell Characterization, Control

Journal: Antioxidants

Article Title: Differential Effects of Three Medium-Chain Fatty Acids on Mitochondrial Quality Control and Skeletal Muscle Maturation

doi: 10.3390/antiox13070821

Figure Lengend Snippet: Effects of C8-induced autophagy enhancement on mitochondrial turnover. C2C12 cells were induced to myotube differentiation for 48 h with pretreatment with C8 (48 h). We then investigated the effects of C8 on myotube mitochondria when autophagy was inhibited using chloroquine (CQ). ( A ) Mitophagy fluorescence staining. ( B ) Semi-quantification of panel ( A ). ( C ) Changes in mitochondria-related protein levels. ( D ) Semi-quantification of panel ( C ). ( E ) Flux analysis. ( F ) Energy metabolic parameters calculated from the flux analysis. ( G ) Mitochondrial superoxide levels. ( H ) Semi-quantitative analysis of panel G. Scale bar, 50 μm; error bar, standard deviation from three independent trials. Statistical differences were calculated using analysis of variance (ANOVA) with Bonferroni correction. C8, caprylic acid; CQ, chloroquine; MYH, myosin heavy chain; CTRL, control; C8, caprylic acid; PGC1α, peroxisome proliferator-activated receptor gamma coactivator 1-α; TOM20, translocase of the outer mitochondrial membrane 20; DRP1, dynamin-related protein 1; MFN2, mitofusin 2; PINK1, PTEN-induced kinase 1; LC3, microtubule-associated protein 1A/1B-light chain 3.

Article Snippet: C2C12 myoblasts were pretreated with C8 (200 μg/mL) and chloroquine (CQ, 10 μM, HY-17589, Med Chem Express, Princeton, NJ, USA) for 48 h and then treated using the differentiation procedure described above.

Techniques: Fluorescence, Staining, Standard Deviation, Control, Membrane