Bnip3ΔTM does not affect rotenone-stimulated autophagic flux.

<p>Representative images of LAMP2 stained WT and PINK1-/- myocytes overexpressing β-gal or BNIP3ΔTM plus MitoGFP after rotenone treatment (90 min).</p

Rotenone induces delayed Parkin translocation to mitochondria in PINK1-/- myocytes.

<p>(A) The mitochondrial complex I inhibitor rotenone (40 μM) significantly reduced mitochondrial membrane potential by 1 hour of treatment in WT and PINK1-/- cardiac myocytes (n = 3). (B) Quantitation of percentage of cells with Parkin translocation to mitochondria after 60 and 90 min of rotenone treatment. (C) Representative images. Mean ± SEM, n = 3, *p<0.05, **p<0.01 vs. 0 min. n.s. = not significant</p

The dominant negative BNIP3ΔTM has no affect rotenone-mediated mitophagy.

<p>(A) Representative fluorescent images of WT and PINK1-/- adult mouse cardiac myocytes overexpressing β-gal or BNIP3ΔTM plus GFP-LC3. (B) Quantitation of the number of autophagosomes in WT and PINK1-/- myocytes. (C) Quantitation of GFP-LC3 puncta colocalization with mitochondrial TOM20. BNIP3ΔTM did not reduce rotenone-mediated mitophagy in either WT or PINK1-/- myocytes. Mean ± SEM, n = 3. n.s. = not significant</p

Rotenone induces activation of autophagy and mitophagy in WT and PINK1-/- adult mouse myocytes.

<p>(A) Quantitation of autophagy activation after 60 and 90 min rotenone (40 μM) treatment in WT and PINK1-/- myocytes. (B) Representative fluorescent images of WT and PINK1-/- myocytes overexpressing GFP-LC3 and stained with anti-TOM20 to label mitochondria. Co-localization between GFP-LC3 and TOM20 indicate activation of mitophagy. (C) Quantitation of the number of GFP-LC3 puncta in cells. (D) Quantitation of the number of GFP-LC3 autophagosomes co-localizing with TOM20-labeled mitochondria. (E) Transmission electron microscopy of isolated adult mouse myocytes after treatment with DMSO or 40 μM rotenone. Autophagosomes containing mitochondria were identified in both WT and PINK1-/- cardiac myocytes after rotenone treatment. Mean ± SEM, n = 3. n.s. = not significant</p

BNIP3 induces mitophagy in WT and PINK1-/- myocytes.

<p>(A) Representative fluorescent images of WT and PINK1-/- adult mouse cardiac myocytes overexpressing GFP-LC3 and either β-gal or BNIP3. (B) The percentage of cells with activated autophagy significantly increased in both WT and PINK1-/- cells in response to BNIP3 overexpression. (C) Quantitation of GFP-LC3 puncta in adult mouse cardiac myocytes show no difference in autophagosome number in PINK1-/- cells compared to WT. (D) Quantitation of GFP-LC3 puncta colocalization with mitochondrial TOM20 show increased mitophagy in both WT and PINK1-/- myocytes in response to BNIP3. (E) Representative images of LAMP2 stained WT and PINK1-/- myocytes overexpressing β-gal or BNIP3. (F) Quantitation of WT and PINK1-/- myocytes with enhanced lysosomal activity. Mean ± SEM, n = 3–4.</p

Autophagic flux is intact in rotenone treated PINK1-/- myocytes.

<p>(A) Representative images of WT and PINK1-/- myocytes overexpressing MitoGFP and stained with anti-LAMP2 after 90 min of rotenone treatment. (B) Quantitation of WT and PINK1-/- myocytes with enhanced lysosomal activity. Mean ± SEM, n = 3.</p

A Unique Non-catenane Interlocked Self-Assembled Supramolecular Architecture and Its Photophysical Properties

A novel, interlocked, self-assembled (M₂L₂)₂ molecular architecture was constructed from an arene-Ru acceptor and a 1,4-di(pyridin-4-yl)buta-1,3-diyne donor. Two M₂L₂ units, with cavities of ∼7.21 Å, spontaneously interlock, with one unit encapsulating a twin in a non-catenane fashion. The dimeric host–guest complex thus formed is unique among two-dimensional self-assemblies and is stabilized by π–π interactions between the M₂L₂ units

Ru(II)-Catalyzed Selective C–H Amination of Xanthones and Chromones with Sulfonyl Azides: Synthesis and Anticancer Evaluation

A ketone-assisted ruthenium-catalyzed selective amination of xanthones and chromones C–H bonds with sulfonyl azides is described. The reactions proceed efficiently with a broad range of substrates with excellent functional group compatibility. This protocol provides direct access to 1-aminoxanthones, 5-aminochromones, and 5-aminoflavonoid derivatives known to exhibit potent anticancer activity