Journal: Frontiers in pharmacology

Article Title: Number 2 Feibi Recipe Inhibits H 2 O 2 -Mediated Oxidative Stress Damage of Alveolar Epithelial Cells by Regulating the Balance of Mitophagy/Apoptosis.

doi: 10.3389/fphar.2022.830554

Figure Lengend Snippet: FIGURE 7 | FBR-2 promotes mitophagy. (A) Fluorescence colocalization between mitochondria (MTR) and lysosomes (LTG). (B) PINK1 mRNA levels (n = 6). (C) Parkin mRNA levels (n = 6). (D) LC3B mRNA levels (n = 6). (E) The blots of PINK1, Parkin, LC3B and β-actin. (F) PINK1 protein levels (n = 3). (G) Parkin protein levels (n = 3). (H) The ratio of LC3B II/I (n = 3). **p < 0.01, compared with the control group; #p < 0.05 and ##p < 0.01, compared with the H2O2 group.

Article Snippet: Nonspecific binding was blocked with 5% nonfat milk for 1 h and then incubated overnight at 4°C with antibodies against PINK1 (1: 500, Cat No. 23274-1-AP, Proteintech), Parkin (1: 500, Cat No. 14060-1-AP, Proteintech), LC3B (1: 1,000, Cat No. ab48394, Abcam), HK-2 (1: 2000, Cat No. 22029-1-AP, Proteintech), XIAP (1: 500, Cat No. 10037-1- Ig, Proteintech), Bcl-2 (1: 1,000, Cat No. ab196495, Abcam), Bax (1: 4,000, Cat No. 50599-2-Ig, Proteintech), and β-actin (1: 50,000, AC026, Abclonal).

Techniques: Fluorescence, Control

Journal: The Journal of Biological Chemistry

Article Title: The nuclear receptor RORα preserves cardiomyocyte mitochondrial function by regulating caveolin-3-mediated mitophagy

doi: 10.1016/j.jbc.2021.101358

Figure Lengend Snippet: Hypoxia-induced mitophagy is blunted by the absence of RORα in vivo and in vitro . A , immunoblotting of mitochondrial fraction from WT and RORα sg/sg heart lysates. B , quantitative RT-PCR on hearts of mice exposed to normoxia or hypoxia (10% O 2 ) for 24 h, normalized to Actb and Tbp . C , immunoblotting of heart lysates of mice exposed to normoxia or hypoxia. D , immunofluorescence staining of normoxic or hypoxic (1% O 2 for 24 h) H9c2 cardiomyoblasts exposed to vehicle, RORα agonist SR1078 (10 mM), or RORα inverse agonist SR3335 (20 mM). E , quantitative analysis of phagolysosomes as identified by costaining LC3B, the lysosomal stain LysoTracker, and mitochondria using ImageJ. F , immunoblotting of NRVM lysates exposed to hypoxia in the presence or absence of bafilomycin A with SR3335 (20 μM) or ( G ) SR1078 (10 μM) with summary densitometry (ImageJ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by unpaired t test ( A – C ) or one-way ANOVA with Tukey's post hoc test ( E – G ). BFA, bafilomycin A; Bnip3, BCL2 interacting protein 3; Hif1a, hypoxia-inducible factor1 alpha; LC3, microtubule-associated protein 1A/1B light chain 3; Pink1, PTEN-induced kinase 1; VDAC1, voltage-dependent anion channel 1.

Article Snippet: RORα (GTX100029, 1:1000), SQSTM1/p62 (GTX629890, 1:1000), PINK1 (GTX107851, 1:1000), LC3B (GTX127375, 1:1000), Ambra1 (GTX55507, 1:1000), and HIF1α (GTX628480) were from Genetex.

Techniques: In Vivo, In Vitro, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: The Journal of Biological Chemistry

Article Title: The nuclear receptor RORα preserves cardiomyocyte mitochondrial function by regulating caveolin-3-mediated mitophagy

doi: 10.1016/j.jbc.2021.101358

Figure Lengend Snippet: Mitophagy is limited in cardiomyocyte-specific RORα KO mouse hearts. A , quantitative RT-PCR (qRT-PCR) for Cre, RORα, and RORγ genes in the heart, liver, and kidney from CMWT and cardiomyocyte-specific RORα KO mice (n = 3 per group). B , qRT-PCR for RORα in the heart tissue from CMWT and CMKO mice exposed to normoxia (21% O 2 ) or to hypoxia (10% O 2 ) for 8 h. C , immunoblotting from WT and CMKO heart lysates with summary densitometry (ImageJ). D , representative LC3B immunofluorescence images of mouse heart sections (≥3 per mouse) and ImageJ summary quantification. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 by unpaired t test ( A ) or one-way ANOVA with Tukey's post hoc test ( B – D ). Atg7, autophagy related 7; Atg10, autophagy related 10; Bnip3, BCL2 interacting protein 3; Cre, Cre recombinase; DAPI, 4′,6-diamidino-2-phenylindole; Hif1a, hypoxia inducible factor1 alpha; Hsp60, heat shock protein 60; LC3B, microtubule-associated protein 1A/1B light chain 3; PINK1, PTEN-induced kinase 1; p62, SQSTM1.

Article Snippet: RORα (GTX100029, 1:1000), SQSTM1/p62 (GTX629890, 1:1000), PINK1 (GTX107851, 1:1000), LC3B (GTX127375, 1:1000), Ambra1 (GTX55507, 1:1000), and HIF1α (GTX628480) were from Genetex.

Techniques: Quantitative RT-PCR, Western Blot, Immunofluorescence