pex16 antibody  (Proteintech)

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: Antibody assay kits and other material sources.

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Acid Assay, Cholesterol Assay, Colorimetric Assay

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced body weight gain in the Pex16 fl/fl mice and Pex16 AdipoQ-Cre mice but not in the Pex16 Alb-Cre mice. ( A ) Body weight gain in the HFD-fed mice (n = 5); ( B ) Repeated Measures ANOVA analysis; ( C ) Liver index (n = 5); ( D ) Fat index (n = 5); ( E ) H&E staining showing adipose inflammation as indicated by “Crown”. The images are representatives of the 5 mice. ( F ) H&E staining showing lipid droplets (Arrows) in liver sections. The images are representatives of the 5 mice. WT, Pex16 fl/fl mice; AKO, adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice; LKO, liver-specific PEX16 knockout ( Pex16 Alb-Cre ) mice.

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Staining, Knock-Out

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced glucose intolerance in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) HFD-induced gonadal adipose tissue expansion as indicated by fat index (n = 5); ( B ) HFD-induced hyperglycemia (n = 5); ( C ) Glucose tolerance test (n = 3).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques:

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced steatosis in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) Liver index (n = 5); ( B ) Liver TG contents (n = 5); ( C ) H&E staining showing lipid droplets (Arrows) in liver sections; ( D ) Steatosis quantification (n = 5); ( E ) Hepatocyte nuclear number (n = 5).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 leads to hepatocyte proliferation. ( A ) PCNA positive staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( B ) PCNA staining quantification (n = 5). ( C ) Ki67 staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( D ) Ki67 staining quantification (n = 5). Arrows show representative positive staining.

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 lowered serum-free fatty acids, ketone bodies and TG. ( A ) Expression of liver peroxisomal fatty acid β-oxidation enzymes; ( B ) Serum fatty acids (n = 5); ( C ) Serum β-hydroxybutyrate (n = 5); ( D ) Expression of fat metabolism enzymes; ( E ) Serum TG (n = 5).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 altered cholesterol and bile acid metabolism. ( A ) Expression of liver cholesterol and bile acid synthetic enzymes; ( B ) Serum cholesterol (n = 5); ( C ) Serum bile acids (n = 5).

Article Snippet: PEX16 antibody , 14816-1-AP , Proteintech, Rosemont, IL, USA.

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: Antibody assay kits and other material sources.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Acid Assay, Cholesterol Assay, Bioassay, Colorimetric Assay, Control

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced body weight gain in the Pex16 fl/fl mice and Pex16 AdipoQ-Cre mice but not in the Pex16 Alb-Cre mice. ( A ) Body weight gain in the HFD-fed mice (n = 5); ( B ) Repeated Measures ANOVA analysis; ( C ) Liver index (n = 5); ( D ) Fat index (n = 5); ( E ) H&E staining showing adipose inflammation as indicated by “Crown”. The images are representatives of the 5 mice. ( F ) H&E staining showing lipid droplets (Arrows) in liver sections. The images are representatives of the 5 mice. WT, Pex16 fl/fl mice; AKO, adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice; LKO, liver-specific PEX16 knockout ( Pex16 Alb-Cre ) mice.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining, Knock-Out

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced glucose intolerance in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) HFD-induced gonadal adipose tissue expansion as indicated by fat index (n = 5); ( B ) HFD-induced hyperglycemia (n = 5); ( C ) Glucose tolerance test (n = 3).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques:

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced steatosis in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) Liver index (n = 5); ( B ) Liver TG contents (n = 5); ( C ) H&E staining showing lipid droplets (Arrows) in liver sections; ( D ) Steatosis quantification (n = 5); ( E ) Hepatocyte nuclear number (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 leads to hepatocyte proliferation. ( A ) PCNA positive staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( B ) PCNA staining quantification (n = 5). ( C ) Ki67 staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( D ) Ki67 staining quantification (n = 5). Arrows show representative positive staining.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 lowered serum-free fatty acids, ketone bodies and TG. ( A ) Expression of liver peroxisomal fatty acid β-oxidation enzymes; ( B ) Serum fatty acids (n = 5); ( C ) Serum β-hydroxybutyrate (n = 5); ( D ) Expression of fat metabolism enzymes; ( E ) Serum TG (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 altered cholesterol and bile acid metabolism. ( A ) Expression of liver cholesterol and bile acid synthetic enzymes; ( B ) Serum cholesterol (n = 5); ( C ) Serum bile acids (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: Antibody assay kits and other material sources.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Acid Assay, Cholesterol Assay, Bioassay, Colorimetric Assay, Control

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced body weight gain in the Pex16 fl/fl mice and Pex16 AdipoQ-Cre mice but not in the Pex16 Alb-Cre mice. ( A ) Body weight gain in the HFD-fed mice (n = 5); ( B ) Repeated Measures ANOVA analysis; ( C ) Liver index (n = 5); ( D ) Fat index (n = 5); ( E ) H&E staining showing adipose inflammation as indicated by “Crown”. The images are representatives of the 5 mice. ( F ) H&E staining showing lipid droplets (Arrows) in liver sections. The images are representatives of the 5 mice. WT, Pex16 fl/fl mice; AKO, adipose-specific PEX16 knockout ( Pex16 AdipoQ-Cre ) mice; LKO, liver-specific PEX16 knockout ( Pex16 Alb-Cre ) mice.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining, Knock-Out

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced glucose intolerance in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) HFD-induced gonadal adipose tissue expansion as indicated by fat index (n = 5); ( B ) HFD-induced hyperglycemia (n = 5); ( C ) Glucose tolerance test (n = 3).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques:

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: HFD-induced steatosis in the Pex16 fl/fl mice but not in the Pex16 Alb-Cre mice. ( A ) Liver index (n = 5); ( B ) Liver TG contents (n = 5); ( C ) H&E staining showing lipid droplets (Arrows) in liver sections; ( D ) Steatosis quantification (n = 5); ( E ) Hepatocyte nuclear number (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 leads to hepatocyte proliferation. ( A ) PCNA positive staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( B ) PCNA staining quantification (n = 5). ( C ) Ki67 staining was observed in the Pex16 Alb-Cre mice, but not in the Pex16 fl/fl mice. Arrows show representative positive staining. The images are representative of the 5 mice. ( D ) Ki67 staining quantification (n = 5). Arrows show representative positive staining.

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Staining

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 lowered serum-free fatty acids, ketone bodies and TG. ( A ) Expression of liver peroxisomal fatty acid β-oxidation enzymes; ( B ) Serum fatty acids (n = 5); ( C ) Serum β-hydroxybutyrate (n = 5); ( D ) Expression of fat metabolism enzymes; ( E ) Serum TG (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing

Journal: Biomedicines

Article Title: Hepatocyte-Specific PEX16 Abrogation in Mice Leads to Hepatocyte Proliferation, Alteration of Hepatic Lipid Metabolism, and Resistance to High-Fat Diet (HFD)-Induced Hepatic Steatosis and Obesity

doi: 10.3390/biomedicines12050988

Figure Lengend Snippet: The absence of liver PEX16 altered cholesterol and bile acid metabolism. ( A ) Expression of liver cholesterol and bile acid synthetic enzymes; ( B ) Serum cholesterol (n = 5); ( C ) Serum bile acids (n = 5).

Article Snippet: Primary antibodies were purchased from international suppliers: ACOX-1 (1:300, Atlas Antibodies, Stockholm, Sweden); PEX16, PMP70, and Catalase (1:500, Bioss Antibodies, Beijing, China); Cyclooxygenase 2 (1:500, Abcam, Boston, MA, USA); MDA (1:100, Kanglang Biotechology, Shanghai, China).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Knockdown of PEX16 Induces Autophagic Degradation of Peroxisomes

doi: 10.3390/ijms22157989

Figure Lengend Snippet: Knockdown of PEX3, PEX16, and PEX19 decreases peroxisome abundance in RPE-1 cells. RPE-1 cells were transfected with siRNA of PEX3, PEX16, and PEX19 for 72 h. The transfection efficiency was evaluated by measuring ( A ) the relative mRNA levels with RT-qPCR and ( B ) the protein expression by immunoblotting. ( C ) Cells were treated with siRNA of PEX3, PEX16, and PEX19 for 48 h and treated with 5 μM chloroquine (CQ) for an additional 24 h. Cells were subjected to immunofluorescence to examine the expression of ABCD3 puncta. ( D ) Quantification of ABCD3 puncta represented the number of peroxisomes per cell. Data are expressed as means ± S.D. ( n = 3, independent experiments), * p < 0.05.

Article Snippet: Reagents and antibodies used were as follows: anti-Calnexin (#ab22595, Abcam, Cambridge, MA, USA), anti-LC3 (#L8918, Sigma-Aldrich, St. Louis, MO, USA), anti-NBR1 (#16004-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX3 (#247042, Abcam), anti-PEX14 (#sc-23197, Santa Cruz Biotechnology, Dallas, TX, USA), anti-PEX16 (#14816-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX19 (#PA5-22129, Invitrogen, Carlsbad, CA, USA), anti-ABCD3 (#ab3421, Abcam, Cambridge, MA, USA and #SAB4200181, Sigma-Aldrich, St. Louis, MO, USA), anti-SQSTM1/p62 (#H00008878-M01, Abnova, Taipei, Taiwan), anti-VDAC (#55259-1-AP, Proteintech, Rosemont, IL, USA), anti-β-actin (#sc-47778, Santa Cruz Biotechnology, Dallas, TX, USA), and chloroquine (#C6628, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Transfection, Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Knockdown of PEX16 Induces Autophagic Degradation of Peroxisomes

doi: 10.3390/ijms22157989

Figure Lengend Snippet: Pexophagy mediates the loss of peroxisomes in RPE-1 cells with PEX16 knockdown. ( A ) Western blot analysis was used on RPE-1 cell lysates to check the protein expression of PEX16, ABCD3, PEX14, Calnexin, and VDAC, using β-actin as an internal control. ( B ) Cells were treated with PEX16 siRNA for 72 h, immunostained with ABCD3, and ( C ) the number of peroxisomes per cell was quantified. Data are expressed as means ± S.D. ( n = 3, independent experiments), * p < 0.05. ( D ) Cells were transfected with PEX16 siRNA, and then treated with chloroquine (CQ) at different time intervals as indicated. ( E ) Then, cells were subjected to immunofluorescence with ABCD3 antibody. ( F ) The number of ABCD3 puncta per cell was quantified. All data are presented as means ± S.D. ( n = 3, independent experiments), * p < 0.05. ( G ) Cells were treated with either control siRNA or PEX16 siRNA for 12 h, incubated with 5 μM CQ for 60 h, and the protein expression of PEX16, LC3, and p62 was examined with western blotting. ( H ) Wild type and ATG5 null MEF cells were treated with PEX16 siRNA for 72 h, and then subjected to immunofluorescence analysis with ABCD3 antibody. ( I ) Quantification of ABCD3 intensity from ( H ). All data are presented as means ± S.D. ( n = 3, independent experiments), * p < 0.05.

Article Snippet: Reagents and antibodies used were as follows: anti-Calnexin (#ab22595, Abcam, Cambridge, MA, USA), anti-LC3 (#L8918, Sigma-Aldrich, St. Louis, MO, USA), anti-NBR1 (#16004-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX3 (#247042, Abcam), anti-PEX14 (#sc-23197, Santa Cruz Biotechnology, Dallas, TX, USA), anti-PEX16 (#14816-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX19 (#PA5-22129, Invitrogen, Carlsbad, CA, USA), anti-ABCD3 (#ab3421, Abcam, Cambridge, MA, USA and #SAB4200181, Sigma-Aldrich, St. Louis, MO, USA), anti-SQSTM1/p62 (#H00008878-M01, Abnova, Taipei, Taiwan), anti-VDAC (#55259-1-AP, Proteintech, Rosemont, IL, USA), anti-β-actin (#sc-47778, Santa Cruz Biotechnology, Dallas, TX, USA), and chloroquine (#C6628, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Western Blot, Expressing, Transfection, Immunofluorescence, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Knockdown of PEX16 Induces Autophagic Degradation of Peroxisomes

doi: 10.3390/ijms22157989

Figure Lengend Snippet: p62 mediates pexophagy in RPE-1 cells with PEX16 knockdown. ( A ) Cells were co-treated with siRNA of NBR1 and PEX16, followed by immunostaining for ABCD3. ( B ) Quantification of ABCD3 puncta per cell from ( A ). ( C ) Cells were co-treated with p62 and PEX16 siRNA and immunostained for ABCD3. ( D ) Quantification of ABCD3 puncta per cell ( C ). Data are expressed as mean ± S.D. ( n = 3, independent experiments), * p < 0.05. ( E ) Cell lysates were also analyzed by western blot to determine the protein expression of PEX16, p62, and ABCD3. Cells were treated with either control siRNA or PEX16 siRNA for 12 h and incubated with 5 μM chloroquine (CQ) for 60 h. ( F ) Co-localization of ABCD3 and NBR1 was analyzed by co-immunostaining. ( G ) Quantification of NBR1/ABCD3 co-localization. ( H ) Co-localization of ABCD3 and p62 was analyzed by co-immunostaining. ( I ) Quantification of p62/ABCD3 co-localization. All data are presented as means ± S.D. ( n = 3–4, independent experiments), * p < 0.05.

Article Snippet: Reagents and antibodies used were as follows: anti-Calnexin (#ab22595, Abcam, Cambridge, MA, USA), anti-LC3 (#L8918, Sigma-Aldrich, St. Louis, MO, USA), anti-NBR1 (#16004-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX3 (#247042, Abcam), anti-PEX14 (#sc-23197, Santa Cruz Biotechnology, Dallas, TX, USA), anti-PEX16 (#14816-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX19 (#PA5-22129, Invitrogen, Carlsbad, CA, USA), anti-ABCD3 (#ab3421, Abcam, Cambridge, MA, USA and #SAB4200181, Sigma-Aldrich, St. Louis, MO, USA), anti-SQSTM1/p62 (#H00008878-M01, Abnova, Taipei, Taiwan), anti-VDAC (#55259-1-AP, Proteintech, Rosemont, IL, USA), anti-β-actin (#sc-47778, Santa Cruz Biotechnology, Dallas, TX, USA), and chloroquine (#C6628, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Immunostaining, Western Blot, Expressing, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Knockdown of PEX16 Induces Autophagic Degradation of Peroxisomes

doi: 10.3390/ijms22157989

Figure Lengend Snippet: Peroxisome function is not recovered by chloroquine in cells with PEX16 knockdown. RPE-1 cells were treated with siRNA of either control or PEX16 for 12 h, followed by incubation with 5 μM chloroquine for 60 h. Under this condition, ( A ) the levels of cholesterol and ( B ) plasmalogens were measured. ( C ) VLCFA (C24:0) level was measured. All data are presented as means ± S.D. ( n = 3, independent experiments), * p < 0.05.

Article Snippet: Reagents and antibodies used were as follows: anti-Calnexin (#ab22595, Abcam, Cambridge, MA, USA), anti-LC3 (#L8918, Sigma-Aldrich, St. Louis, MO, USA), anti-NBR1 (#16004-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX3 (#247042, Abcam), anti-PEX14 (#sc-23197, Santa Cruz Biotechnology, Dallas, TX, USA), anti-PEX16 (#14816-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX19 (#PA5-22129, Invitrogen, Carlsbad, CA, USA), anti-ABCD3 (#ab3421, Abcam, Cambridge, MA, USA and #SAB4200181, Sigma-Aldrich, St. Louis, MO, USA), anti-SQSTM1/p62 (#H00008878-M01, Abnova, Taipei, Taiwan), anti-VDAC (#55259-1-AP, Proteintech, Rosemont, IL, USA), anti-β-actin (#sc-47778, Santa Cruz Biotechnology, Dallas, TX, USA), and chloroquine (#C6628, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Incubation

Journal: International Journal of Molecular Sciences

Article Title: Knockdown of PEX16 Induces Autophagic Degradation of Peroxisomes

doi: 10.3390/ijms22157989

Figure Lengend Snippet: Primer sequences used in the present study.

Article Snippet: Reagents and antibodies used were as follows: anti-Calnexin (#ab22595, Abcam, Cambridge, MA, USA), anti-LC3 (#L8918, Sigma-Aldrich, St. Louis, MO, USA), anti-NBR1 (#16004-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX3 (#247042, Abcam), anti-PEX14 (#sc-23197, Santa Cruz Biotechnology, Dallas, TX, USA), anti-PEX16 (#14816-1-AP, Proteintech, Rosemont, IL, USA), anti-PEX19 (#PA5-22129, Invitrogen, Carlsbad, CA, USA), anti-ABCD3 (#ab3421, Abcam, Cambridge, MA, USA and #SAB4200181, Sigma-Aldrich, St. Louis, MO, USA), anti-SQSTM1/p62 (#H00008878-M01, Abnova, Taipei, Taiwan), anti-VDAC (#55259-1-AP, Proteintech, Rosemont, IL, USA), anti-β-actin (#sc-47778, Santa Cruz Biotechnology, Dallas, TX, USA), and chloroquine (#C6628, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: