Journal: Scientific Reports

Article Title: Mitoxantrone modulates a heparan sulfate-spike complex to inhibit SARS-CoV-2 infection

doi: 10.1038/s41598-022-10293-x

Figure Lengend Snippet: Mitoxantrone inhibits SARS-CoV-2 infection in an EpiAirway 3D tissue model. (A) A schematic diagram of the experimental design. (B) Remdesivir (2 μM) but not Bleomycin (100 μM) inhibits SARS-CoV-2 infection. 24 or 96 h after drug treatment and viral infection (MOI 0.1), the cell surface was washed. The viral titer (TCID50) in the wash was determined. (C,D) Mitoxantrone inhibits SARS-CoV-2 infection in the EpiAirway 3D model. TCID50 was determined either 24 h (C) or 96 h (D) after the organoids were treated with the drug at the indicated concentrations and then air-infected with SARS-CoV-2 at MOI 0.1 for 1 h. The cells were washed from the apical side to remove the virus in the cell exterior and then incubated for 24 (C) or 96 h (D) . Cells were rinsed from the apical side again and viral titers in the wash were determined. The dashed lines indicate the viral titer from cells infected without Mitoxantrone or in the presence of 2 mM Remdesivir (Rem.), as indicated. (E) Bleomycin (100 μM) but not Remdesivir (2 μM) induces cell death, releasing LDH as determined by a luciferase assay. *, p < 0.05, **, p < 0.01, ***, p < 0.001, ****, p < 0.0001 by unpaired student’s t-test. n = 2 tissues per test, each with 3 technical repeats. (F,G) Mitoxantrone inhibits SARS-CoV-2-induced cytotoxicity in the EpiAirway 3D model. As in (D,E) except that LDH in washes collected from Mitoxantrone-treated tissues was measured.

Article Snippet: Medium from the basolateral layer of the tissue culture inserts was removed 24- and 96-h post-infection and diluted in LDH Storage Buffer as per the manufacturer’s instructions (LDH-Glo Cytotoxicity Assay, Promega).

Techniques: Infection, Virus, Incubation, Luciferase

Journal: Bioengineering & Translational Medicine

Article Title: Photobiomodulation augments the effects of mitochondrial transplantation in the treatment of spinal cord injury in rats by facilitating mitochondrial transfer to neurons via Connexin 36

doi: 10.1002/btm2.10473

Figure Lengend Snippet: Combined therapy alleviated mitochondrial‐related neuronal apoptosis. (a) Representative images of the whole spinal cord with MAP2 (red) in left panel pictures. Scale bar: 1 mm. The right panel pictures are enlarged from the left one. Scale bar: 50 μm. Calculated the fluorescence intensity of MAP2 in the right panel pictures by Image J. n = 5. (b) TUNEL (red) and NeuN (green) co‐stained, nuclei stained with DAPI (blue). Scale bar: 50 μm. The statistical ratio of TUNLE‐positive neurons by Image J, Arrows point to representative cells. n = 5. (c) Western blot analysis and quantification of Bax, Bcl2, pro‐caspase 3, and active‐caspase 3 expression levels in each group. n = 5.

Article Snippet: A mitochondrial storage solution (C3609, Beyotime, China) was added to resuspend the mitochondria.

Techniques: Fluorescence, TUNEL Assay, Staining, Western Blot, Expressing

Journal: Bioengineering & Translational Medicine

Article Title: Photobiomodulation augments the effects of mitochondrial transplantation in the treatment of spinal cord injury in rats by facilitating mitochondrial transfer to neurons via Connexin 36

doi: 10.1002/btm2.10473

Figure Lengend Snippet: In the context of mitochondrial transplantation, PBM promoted the expression of Cx36, KIF5B, TRAK2, and Rhot1 and reduced the expression of SNPH. (a) Western blot analysis and quantification of KIF5B, TRAK2, Rhot1, SNPH, and Cx36 expression levels at 1, 3, 7, and 14 dpi in each group. n = 5. (b) Neurons stained with MAP2 (green) co‐localized with Cx36 (red) at 3, 7 dpi in SCI+Mito and SCI+Mito+PBM group. Nuclei were stained with DAPI (blue). Scale bar: 20 μm. n = 3.

Article Snippet: A mitochondrial storage solution (C3609, Beyotime, China) was added to resuspend the mitochondria.

Techniques: Transplantation Assay, Expressing, Western Blot, Staining

Journal: Bioengineering & Translational Medicine

Article Title: Photobiomodulation augments the effects of mitochondrial transplantation in the treatment of spinal cord injury in rats by facilitating mitochondrial transfer to neurons via Connexin 36

doi: 10.1002/btm2.10473

Figure Lengend Snippet: In the context of mitochondrial transplantation, PBM promoted the transfer of exogenous mitochondria into neurons. (a) The distribution of platelet‐derived mitochondria which was stained with Mito Tracker Red in the horizontal and sagittal planes of the injured segment. Nuclei were stained with DAPI (blue). R = Rostral; C = Caudal; D = Dorsal; V = Ventral. Scale bar: 100 μm. (b) Neurons stained with MAP2 (green) co‐localized with mitochondria (Mito Tracker Red) at 1, 3, 7, 14 dpi. Nuclei were stained with DAPI (blue). The ratio of mitochondria entering neurons was calculated. Scale bar: 20 μm. n = 4.

Article Snippet: A mitochondrial storage solution (C3609, Beyotime, China) was added to resuspend the mitochondria.

Techniques: Transplantation Assay, Derivative Assay, Staining

Journal: Bioengineering & Translational Medicine

Article Title: Photobiomodulation augments the effects of mitochondrial transplantation in the treatment of spinal cord injury in rats by facilitating mitochondrial transfer to neurons via Connexin 36

doi: 10.1002/btm2.10473

Figure Lengend Snippet: Combined therapy promoted mitochondrial activity. (a) Representative images of neuronal mitochondria in each group by TEM. Qualitative analysis of the proportion of elongated (>2 μm in size) mitochondria. Representative mitochondria are indicated by red arrows. Left panel scale bar: 2 μm. Right panel scale bar: 1 μm. n = 5. (b) Representative images of mitochondria stained with Tom20 antibody (red) at 14 dpi. Nuclei were stained with DAPI (blue). Images were separated, thresholded, filtered, and binarized with Image J. Continuous mitochondrial structures were calculated as the percentage of the area of large particles normalized to the total mitochondrial particles area. Scale bar: 10 μm. n = 5. (c–h) Western blot analysis and quantification of the expression levels of Complex I‐V in each group. n = 5. (i) Result of ATP levels in each group. n = 5.

Article Snippet: A mitochondrial storage solution (C3609, Beyotime, China) was added to resuspend the mitochondria.

Techniques: Activity Assay, Staining, Western Blot, Expressing