Journal: Oncogene

Article Title: Mitochondrial structure and function adaptation in residual triple negative breast cancer cells surviving chemotherapy treatment

doi: 10.1038/s41388-023-02596-8

Figure Lengend Snippet: (A) Schematic diagram of procedures performed on PDX tumors isolated from NOD/SCID mice bearing orthotopic PIM001-P tumors treated with a single AC dose (day 0; 50 mg/kg C + 0.5 mg/kg A). Tumors were harvested prior to AC treatment, 2 days following AC (ACd2), 21 days following AC (residual), and 35 days following AC when tumors had reached the starting tumor volume (regrown) and were used for multiple analyses: (B) Seahorse MitoStress Tests were conducted immediately after tumor isolation, digestion, and purification of human tumor cells. Maximal OCR, basal OCR, basal ECAR and OCR/ECAR ratio were computed from the MitoStress Test. ****P < 0.0001 and *<0.05 by one-way ANOVA test. (C) IHC with a human mitochondria specific antibody. Scale bars are 20 μm. (D) Vectra 3 machine learning quantification of IHC staining intensity within tumor cells. *P<0.05 by one-way ANOVA test. (E) qPCR-based quantification of mtDNA:nucDNA ratio using DNA prepared from snap-frozen purified tumor cells. (F-G) TEM to quantify mitochondrial length. Mitochondria were annotated with QuPath. Scale bars are 1 μm. Approximately 3000 mitochondria per group were analyzed across 3 biological replicate mice per group. ***P < 0.001 and *<0.05 by one-way ANOVA test, (H) western blotting to measure mitochondrial fusion and fission protein levels, and (I) co-immunofluorescence of OPA1 (red), VDAC1 (green; see Fig S11), and nucleus DAPI (blue). scale bars are 10 μm. Fluorescence intensity of OPA1 normalized to VDAC was measured by ImageJ. ****P < 0.0001, ***<0.001, **<0.01, *<0.05 by one-way ANOVA test.

Article Snippet: Mitochondrial DNA damage assay Long amplification PCR (LA-PCR) was performed in a 25 μl reaction on mixture containing 2.5units LongAmp Taq DNA Polymerase, 1x LongAmp Taq Reaction Buffer, 300 μM of dNTPs (New England Biolabs, E5200S), 0.5 μM of each primer (8kb mitochondrial DNA fragment, forward: 5’-TCTAAGCCTCCTTATTCGAGCCGA-3’, reverse: 5’-TTTCATCATGCGGAGATGTTGGATGG-3’), and 50ng of DNA template.

Techniques: Isolation, Purification, Immunohistochemistry, Western Blot, Immunofluorescence, Fluorescence

Journal: Aquatic toxicology (Amsterdam, Netherlands)

Article Title: Characterization of AHR1 and its Functional Activity in Atlantic Sturgeon and Shortnose Sturgeon

doi: 10.1016/j.aquatox.2018.09.014

Figure Lengend Snippet: Primers used to amplify and sequence Atlantic sturgeon and shortnose sturgeon AHR1

Article Snippet: First strand cDNAs were prepared from Atlantic and shortnose sturgeon as described above and subjected to Extra-Long PCR amplification using a LongAmp® Taq PCR Kit (New England Biolabs, Ipswich, MA) to separately amplify the 5’ and 3’ halves of both AHR1 and AHR2 coding sequences.

Techniques: Sequencing, Real-time Polymerase Chain Reaction