rabbit polyclonal anti pgam5 antibody  (ProSci Incorporated)

Journal: World Journal of Clinical Cases

Article Title: Clinical and prognostic significance of expression of phosphoglycerate mutase family member 5 and Parkin in advanced colorectal cancer

doi: 10.12998/wjcc.v10.i14.4368

Figure Lengend Snippet: Representative immunohistochemical staining for phosphoglycerate mutase family member 5 and Parkin protein (original magnification 200 ×). The expression levels of phosphoglycerate mutase family member 5 and Parkin protein were scored as negative (0), weak (1), moderate (2), or strong (3) according to staining intensity. PGAM5: Phosphoglycerate mutase family member 5.

Article Snippet: Non-specific binding was blocked using 5% bovine serum albumin in Tris Buffered Saline with Tween 20 (TBS-T, pH 7.6) for 2 h. Subsequently, incubation with primary antibodies, rabbit polyclonal anti-PGAM5 (1:1000 dilution, ab126534; Abcam, United Kingdom), rabbit polyclonal anti-Parkin (1:1000 dilution, ab233434; Abcam, United Kingdom), and glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, 1:1000 dilution, FL-335; Santa Cruz Biotechnology, CA, United States) was undertaken overnight at 4 °C.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: World Journal of Clinical Cases

Article Title: Clinical and prognostic significance of expression of phosphoglycerate mutase family member 5 and Parkin in advanced colorectal cancer

doi: 10.12998/wjcc.v10.i14.4368

Figure Lengend Snippet: Representative immunohistochemical staining for phosphoglycerate mutase family member 5 and Parkin in colorectal adenocarcinoma tissues and adjacent tissues (original magnification 200 ×). Phosphoglycerate mutase family member 5 (PGAM5) and Parkin protein were expressed in the cytoplasm of colonic epithelial cells. Specimens were categorized as either high or low expression according to the Youden index. High expression of PGAM5 was defined as a score of ≥ 4.75, and Parkin was ≥ 7.1. Low expression of PGAM5 was defined as a score of < 4.75, and Parkin was < 7.1. PGAM5: Phosphoglycerate mutase family member 5.

Article Snippet: Non-specific binding was blocked using 5% bovine serum albumin in Tris Buffered Saline with Tween 20 (TBS-T, pH 7.6) for 2 h. Subsequently, incubation with primary antibodies, rabbit polyclonal anti-PGAM5 (1:1000 dilution, ab126534; Abcam, United Kingdom), rabbit polyclonal anti-Parkin (1:1000 dilution, ab233434; Abcam, United Kingdom), and glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, 1:1000 dilution, FL-335; Santa Cruz Biotechnology, CA, United States) was undertaken overnight at 4 °C.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: World Journal of Clinical Cases

Article Title: Clinical and prognostic significance of expression of phosphoglycerate mutase family member 5 and Parkin in advanced colorectal cancer

doi: 10.12998/wjcc.v10.i14.4368

Figure Lengend Snippet: Western blots showing higher phosphoglycerate mutase family member 5 and Parkin levels in colorectal adenocarcinoma fresh tissues than in corresponding paracancerous tissues in six matched pairs (T, colorectal adenocarcinoma tissues; N, adjacent non-tumor tissues). PGAM5: Phosphoglycerate mutase family member 5; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Non-specific binding was blocked using 5% bovine serum albumin in Tris Buffered Saline with Tween 20 (TBS-T, pH 7.6) for 2 h. Subsequently, incubation with primary antibodies, rabbit polyclonal anti-PGAM5 (1:1000 dilution, ab126534; Abcam, United Kingdom), rabbit polyclonal anti-Parkin (1:1000 dilution, ab233434; Abcam, United Kingdom), and glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, 1:1000 dilution, FL-335; Santa Cruz Biotechnology, CA, United States) was undertaken overnight at 4 °C.

Techniques: Western Blot

Journal: World Journal of Clinical Cases

Article Title: Clinical and prognostic significance of expression of phosphoglycerate mutase family member 5 and Parkin in advanced colorectal cancer

doi: 10.12998/wjcc.v10.i14.4368

Figure Lengend Snippet: Kaplan-Meier survival curves for phosphoglycerate mutase family member 5 and Parkin expression in colorectal cancer tissues. PGAM5: Phosphoglycerate mutase family member 5.

Article Snippet: Non-specific binding was blocked using 5% bovine serum albumin in Tris Buffered Saline with Tween 20 (TBS-T, pH 7.6) for 2 h. Subsequently, incubation with primary antibodies, rabbit polyclonal anti-PGAM5 (1:1000 dilution, ab126534; Abcam, United Kingdom), rabbit polyclonal anti-Parkin (1:1000 dilution, ab233434; Abcam, United Kingdom), and glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, 1:1000 dilution, FL-335; Santa Cruz Biotechnology, CA, United States) was undertaken overnight at 4 °C.

Techniques: Expressing

Journal: World Journal of Clinical Cases

Article Title: Clinical and prognostic significance of expression of phosphoglycerate mutase family member 5 and Parkin in advanced colorectal cancer

doi: 10.12998/wjcc.v10.i14.4368

Figure Lengend Snippet: Univariate analysis of overall survival and progression-free survival for colorectal cancer by Cox regression

Article Snippet: Non-specific binding was blocked using 5% bovine serum albumin in Tris Buffered Saline with Tween 20 (TBS-T, pH 7.6) for 2 h. Subsequently, incubation with primary antibodies, rabbit polyclonal anti-PGAM5 (1:1000 dilution, ab126534; Abcam, United Kingdom), rabbit polyclonal anti-Parkin (1:1000 dilution, ab233434; Abcam, United Kingdom), and glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, 1:1000 dilution, FL-335; Santa Cruz Biotechnology, CA, United States) was undertaken overnight at 4 °C.

Techniques:

Journal: World Journal of Clinical Cases

Article Title: Clinical and prognostic significance of expression of phosphoglycerate mutase family member 5 and Parkin in advanced colorectal cancer

doi: 10.12998/wjcc.v10.i14.4368

Figure Lengend Snippet: Multivariate analysis of overall survival and progression-free survival for colorectal cancer by Cox regression

Article Snippet: Non-specific binding was blocked using 5% bovine serum albumin in Tris Buffered Saline with Tween 20 (TBS-T, pH 7.6) for 2 h. Subsequently, incubation with primary antibodies, rabbit polyclonal anti-PGAM5 (1:1000 dilution, ab126534; Abcam, United Kingdom), rabbit polyclonal anti-Parkin (1:1000 dilution, ab233434; Abcam, United Kingdom), and glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate dehydrogenase, 1:1000 dilution, FL-335; Santa Cruz Biotechnology, CA, United States) was undertaken overnight at 4 °C.

Techniques:

Journal: bioRxiv

Article Title: Functional single cell proteomic profiling of cells with abnormal DNA damage response dynamics

doi: 10.1101/2021.10.13.464241

Figure Lengend Snippet: (a) Phototagging of the Group 2 cells. The zoomed-in images showed a representative cell before and after phototagging. Red, 53BP1; green, phototagging (PT) dye. (b) Schematic protocol of single cell proteomics analysis. Four non-tagged cells, four tagged cells and 200 nontagged cells (serving as carrier cells), were labeled with respective 10-plex TMT labels as indicated, and then mixed into one sample before subjecting to LC-MS/MS. 10 samples, in total 40 Group 1 and 40 Group 2 cells, were analyzed. (c) (i) A volcano plot showing proteins enriched either in Group 1 (normal DDR) or Group 2 (abnormal DDR) cells. 80 cells were pooled into the analysis. Dashed red line shows the cutoff of p value at 0.05. (ii) PDS5A and PGAM5 were found upregulated in Group 2 cells and were highlighted. (d) Immunofluorescence quantification of (i) PDS5A and (ii) PGAM5 proteins in Group 1 and Group 2 cells; N = 16 (for PDS5A) and N = 20 (for PGAM5) with p value of 0.0010 and 0.0362, respectively.

Article Snippet: The cells were then incubated with 1:500 rabbit polyclonal anti-PDS5A antibody (Novus), 1:1000 rabbit polyclonal anti-PGAM5 antibody (Novus) or 1:1000 rabbit polyclonal anti-γH2AX antibody (Abcam) for 90 min at room temperature.

Techniques: Labeling, Liquid Chromatography with Mass Spectroscopy, Immunofluorescence

Journal: PLoS Genetics

Article Title: The Loss of PGAM5 Suppresses the Mitochondrial Degeneration Caused by Inactivation of PINK1 in Drosophila

doi: 10.1371/journal.pgen.1001229

Figure Lengend Snippet: (A) Silver-stained polyacrylamide gel to visualize hPINK1-binding proteins. FLAG elution fractions purified from cells stably expressing hPINK1-FLAG (PINK1-FLAG lane) and parental cells (Control lane) are separated on a gel (For details of the procedure, see ). Bands corresponding to hPINK1 (dots) and representative co-purified proteins are indicated. (B, C) The wing phenotype typical of 10- and 20-day-old dPINK1 RNAi flies (B) was suppressed by the PGAM5 NP0568 mutant allele, whereas viability of 10-, 20- and 30-day-old adult flies was not improved (C). *, p <0.05; **, p <0.01 vs. age-matched dPINK1 RNAi group in Student's t -test. The genotypes are as follows: MHC-GAL4> dPINK1 RNAi (+/+), PGAM5 NP0568 /Y; MHC-GAL4> dPINK1 RNAi ( PGAM5 NP0568 /Y ), PGAM5 NP0568 /+; MHC-GAL4> dPINK1 RNAi ( PGAM5 NP0568 /+ ). MHC-GAL4 , a muscle-specific driver. Flies were raised at 29°C as the RNAi-induced dPINK1 defects are more pronounced when flies are raised at that temperature. (D, E) Removal of one copy of the IRS4 ortholog chico had no effect on the wing phenotype of dPINK1 RNAi flies (D) but improved viability (E). *, p <0.05; **, p <0.01 vs. age-matched dPINK1 RNAi group. The genotypes are: MHC-GAL4> dPINK1 RNAi (+/+), chico /+; MHC-GAL4> dPINK1 RNAi ( chico ). Flies were raised at 29°C.

Article Snippet: Rabbit anti-human PGAM5 polyclonal antibody was raised against recombinant GST-tagged PGAM5 domain (89–289 aa) produced in the E. coli strain BL21(DE3)pLysS (Novagen), and was affinity-purified against the antigen.

Techniques: Staining, Binding Assay, Purification, Stable Transfection, Expressing, Mutagenesis

Journal: PLoS Genetics

Article Title: The Loss of PGAM5 Suppresses the Mitochondrial Degeneration Caused by Inactivation of PINK1 in Drosophila

doi: 10.1371/journal.pgen.1001229

Figure Lengend Snippet: (A) Loss of dPGAM5 genes extends the lifespan. Adult male wild-type ( yw/Y ; n = 125) vs. dPGAM5 null ( y , PGAM5 1 /Y; n = 125) flies, p <0.001 by log rank test. (B) Overexpression of dPGAM5 or dPGAM5-2 in Drosophila causes shorter lifespan. Overexpression of the transgenes was induced using the ubiquitous daughterless ( Da ) -GAL4 driver. Lifespan of adult male EGFP ( n = 130), dPGAM5 ( n = 76) and dPGAM5-2 ( n = 117) flies. EGFP vs. dPGAM5 , p <0.001; EGFP vs. dPGAM5-2 , p <0.001; by log rank test. (C–J) Transmission electron microscopy (TEM) analysis of the indirect flight muscle and morphology of mitochondria in 2-day-old adult flies with the indicated genotypes. In C–F, we outlined some mitochondria with broken lines to highlight morphology. The insets in G–J show representative mitochondria matrixes. A revertant, PINK1 RV , was used as a wild-type comparison . The genotypes are: PINK1 RV /Y (C, G), PGAM5 NP0568 /Y (D, H), Da-GAL4> UAS - dPGAM5 (E, I), Da-GAL4> UAS - dPGAM5-2 (F, J). Scale bars = 1 µm in C–F and 200 nm in G–J. (K) Quantification of the percentage of mitochondrial size distribution in the indirect muscle tissue from wild-type ( n = 136 from 5 adult flies), PGAM5 NP0568 ( n = 155 from 5), PGAM5 1 ( n = 87 from 5), dPGAM5 Tg ( n = 143 from 5) and dPGAM5-2 Tg flies ( n = 147 from 5) as shown in (C–J). The length of the mitochondria in the direction of the myofibrils was measured. Data are shown as means ± SE (* p <0.05, ** p <0.01 vs. wild-type ). (L–O) Brain tissues of 5-day-old adult flies were stained with anti-TH antibody (red). Mitochondria labeled with mitoGFP (green) were observed in the PPL1 TH-positive neurons of the indicated genotypes. The genotypes are as follows: TH-GAL4> mitoGFP ( wild-type ), PGAM5 1 /Y; TH-GAL4>UAS-mitoGFP ( PGAM5 1 ), UAS-dPGAM5/TH-GAL4> UAS-mitoGFP ( dPGAM5 Tg ), UAS-dPGAM5-2/TH-GAL4> UAS-mitoGFP ( dPGAM5-2 Tg ). TH-GAL4 , a DA neuron-specific driver. Scale bar = 5 µm.

Article Snippet: Rabbit anti-human PGAM5 polyclonal antibody was raised against recombinant GST-tagged PGAM5 domain (89–289 aa) produced in the E. coli strain BL21(DE3)pLysS (Novagen), and was affinity-purified against the antigen.

Techniques: Over Expression, Transmission Assay, Electron Microscopy, Staining, Labeling

Journal: PLoS Genetics

Article Title: The Loss of PGAM5 Suppresses the Mitochondrial Degeneration Caused by Inactivation of PINK1 in Drosophila

doi: 10.1371/journal.pgen.1001229

Figure Lengend Snippet: (A–F) dPGAM5 inactivation failed to rescue the mitochondrial fragmentation caused by mfn knockdown ( mfn RNAi ) or introduction of an extra copy of the drp1 gene ( drp1 + ). To visualize the mitochondria under a fluorescence microscopy, we used the muscle-specific MHC-GAL4 driver to induce expression of a mitoGFP (green) transgene in 5-day-old adult flies with the indicated genotypes. Muscle tissue was counterstained with phalloidin (red). Scale bar = 2 µm. (G) The average length of the mitochondria in the direction of the myofibrils was measured from wild-type ( n = 343 from 7 adult flies), PGAM5 1 ( n = 390 from 8), mfn RNAi (VDRC40478, n = 305 from 6; VDRC105261, n = 372 from 8), mfn RNAi (VDRC40478) ; PGAM5 1 ( n = 355 from 7), mfn RNAi (VDRC105261) ; PGAM5 1 ( n = 237 from 5), drp1 + ( n = 245 from 5) and drp1 + ; PGAM5 1 ( n = 247 from 5) as shown in (A–F). Data are shown as means ± SE (** p <0.01; N.S., not significant). The genotypes are as follows: +/Y; MHC-GAL4>mitoGFP (A, wild-type ), PGAM5 1 /Y; MHC-GAL4>UAS-mitoGFP (B, PGAM5 1 ), +/Y; MHC-GAL4>UAS-mitoGFP; UAS-mfn RNAi (VDRC40478) (C, mfn RNAi ), PGAM5 1 /Y; MHC-GAL4>UAS-mitoGFP; UAS-mfn RNAi (VDRC40478) (D, mfn RNAi; PGAM5 1 ), +/Y; MHC-GAL4>mitoGFP; drp1 + (E, drp1 + ), PGAM5 1 /Y; MHC-GAL4>mitoGFP; drp1 + (F, drp1 + ; PGAM5 1 ).

Article Snippet: Rabbit anti-human PGAM5 polyclonal antibody was raised against recombinant GST-tagged PGAM5 domain (89–289 aa) produced in the E. coli strain BL21(DE3)pLysS (Novagen), and was affinity-purified against the antigen.

Techniques: Fluorescence, Microscopy, Expressing

Journal: PLoS Genetics

Article Title: The Loss of PGAM5 Suppresses the Mitochondrial Degeneration Caused by Inactivation of PINK1 in Drosophila

doi: 10.1371/journal.pgen.1001229

Figure Lengend Snippet: A thorax defect (B, arrowheads) and abnormal wing posture (C) caused by loss of dPINK1 activity are suppressed in dPGAM5 mutant genetic backgrounds (A, D and E). (F) Percentage of 10-, 20- and 30-day-old male flies showing abnormal wing postures. Error bars show S.E. from three experiments. (G) Percentage of 10-day-old male PINK1 B9 and PINK1 B9 ubiquitously overexpressing dPGAM5 flies showing abnormal wing postures. Error bars show S.E. from three experiments. (H, I) Percentage of locomotor activity. Error bars show S.E. from three repeated experiments. (J) Lifespan of adult male flies. Loss of dPGAM5 partially improved the reduced lifespan seen in PINK1 B9 fly ( PINK1 B9 vs. PINK1 B9 , PGAM5 NP0568 or PINK1 B9 , PGAM5 1 , p <0.001; wild-type vs. PINK1 B9 , PGAM5 NP0568 or PINK1 B9 , PGAM5 1 , p <0.01 by the log rank test). (K) Lifespan of adult male PINK1 B9 and PINK1 B9 ubiquitously overexpressing dPGAM5 flies. Overexpression of dPGAM5 further reduced the lifespan ( PINK1 B9 vs. PINK1 B9 ; dPGAM5 Tg , p <0.001). The same files were used in (A–F, H and J) and in (G, I and K). The genotypes and the number used in the assays are; wild-type ( PINK1 RV / Y , n = 161), PGAM5 NP0568 ( PGAM5 NP0568 / Y , n = 161), PGAM5 1 ( PGAM5 1 / Y , n = 161), PINK1 B9 ( PINK1 B9 / Y , n = 101), PINK1 B9 , PGAM5 NP0568 ( PINK1 B9 , PGAM5 NP0568 / Y , n = 162) and PINK1 B9 , PGAM5 1 ( PINK1 B9 , PGAM5 1 / Y , n = 160) in (A–F, H and J), PINK1 B9 ( PINK1 B9 / Y; Da-GAL4/+ , n = 162) and PINK1 B9 , dPGAM5 Tg ( PINK1 B9 / Y; Da-GAL4> UAS-dPGAM5 , n = 161) in (G, I and K).

Article Snippet: Rabbit anti-human PGAM5 polyclonal antibody was raised against recombinant GST-tagged PGAM5 domain (89–289 aa) produced in the E. coli strain BL21(DE3)pLysS (Novagen), and was affinity-purified against the antigen.

Techniques: Activity Assay, Mutagenesis, Over Expression

Journal: PLoS Genetics

Article Title: The Loss of PGAM5 Suppresses the Mitochondrial Degeneration Caused by Inactivation of PINK1 in Drosophila

doi: 10.1371/journal.pgen.1001229

Figure Lengend Snippet: (A–F) TEM analysis of the indirect flight muscle and morphology of mitochondria in 2-day-old adult flies with the indicated genotypes. In A and B, some mitochondria are outlined with broken lines. The insets in D–F show representative mitochondria matrixes. Scale bars = 1 µm in A–C and 200 nm in D–F. (G) Quantification of the percentage of mitochondrial size distribution in the indirect muscle tissue from wild-type ( n = 136 from 5 adult flies), PINK1 B9 ( n = 96 from 5), PINK1 B9 , PGAM5 NP0568 ( n = 116 from 5), PINK1 B9 , PGAM5 1 ( n = 111 from 5) as shown in . Data are shown as means ± SE (* p <0.05, ** p <0.01). (H–J) Quantification of the percentage of cytoplasmic mitochondrial aggregates with diameter of 0.5–1.0, 1.0–1.5 or ≥1.5 µm in each PPL1 TH + neuron from wild-type ( n = 373 from 18 adult flies), PGAM5 1 ( n = 356 from 18), PINK1 B9 ( n = 231 from 11), PINK1 B9 PGAM5 1 flies ( n = 235 from 13). Mitochondrial morphology was revealed by mitoGFP as shown in . Data are shown as means ± SE (*, p <0.05;**, p <0.01; N.S., not significant). Tubular or reticular mitochondria were excluded from the estimation due to difficulty in the counting. However, the ratio of mitochondria with that morphology was also increased in PINK1 B9 PGAM5 1 flies (J) compared with that in PINK1 B9 flies (I). Arrowheads in (J) indicate representative tubular or reticular mitochondria. Scale bar in (I) = 5 µm. (K) Quantification of TH + DA neuron number in the PPM1, PPM2 and PPL1 clusters in 25-day-old males. PPM1 and PPM2 cluster neurons were counted together. Data are shown as means ± SE (*, p <0.05; n = 16). (L, M) Representative images of PPM1/2, PPM3 and PPL1 clusters of PINK1 B9 (L) and PINK1 B9 , PGAM5 NP0568 flies (M) visualized with anti-TH antibody. Scale bar in (M) = 50 µm. The genotypes are: PINK1 RV /Y ( wild-type ), PINK1 B9 /Y ( PINK1 B9 ), PGAM5 NP0568 /Y ( PGAM5 NP0568 ), PGAM5 1 /Y ( PGAM5 1 ), PINK1 B9 , PGAM5 NP0568 /Y ( PINK1 B9 , PGAM5 NP0568 ), PINK1 B9 , PGAM5 1 /Y ( PINK1 B9 , PGAM5 1 ), PINK1 B9 /Y ; Da-GAL4 > dPGAM5 ( PINK1 B9 , PGAM5 Tg ), in (A–G, K–M), PINK1 RV /Y; TH-GAL4> mitoGFP ( wild-type ), PINK1 B9 /Y; TH-GAL4> mitoGFP ( PINK1 B9 ), PGAM5 1 /Y; TH-GAL4> mitoGFP ( PGAM5 1 ), PINK1 B9 , PGAM5 1 /Y; TH-GAL4> mitoGFP ( PINK1 B9 , PGAM5 1 ) in (H–J).

Article Snippet: Rabbit anti-human PGAM5 polyclonal antibody was raised against recombinant GST-tagged PGAM5 domain (89–289 aa) produced in the E. coli strain BL21(DE3)pLysS (Novagen), and was affinity-purified against the antigen.

Techniques:

Journal: PLoS Genetics

Article Title: The Loss of PGAM5 Suppresses the Mitochondrial Degeneration Caused by Inactivation of PINK1 in Drosophila

doi: 10.1371/journal.pgen.1001229

Figure Lengend Snippet: The abnormal wing posture caused by a homozygous dParkin mutation (A) was not suppressed by removal of the dPGAM5 gene (B). (C) Percentage of flies with abnormal wing posture among 10- and 20-day-old male wild-type ( n = 105), parkin P21 ( n = 102) and PGAM5 NP0568 ; parkin P21 ( n = 109) flies. Error bars show S.E. from three repeated experiments. *, p <0.05; **, p <0.01 vs. dParkin (+/+). (D) Percentage of flies showing locomotor activity among 10- and 20-day-old male parkin P21 ( n = 86), parkin P21 ( n = 73) and PGAM5 NP0568 ; parkin P21 ( n = 78) flies. Error bars show S.E. from twenty repeated experiments. *, p <0.05; **, p <0.01 vs. dParkin (+/+). (E) Locomotor activity of 10-day-old male parkin P21 ( n = 153) and parkin P21 ubiquitously overexpressing dPGAM5 flies ( parkin P21 ; dPGAM5 Tg , n = 155) flies. Error bars show S.E. from twenty repeated experiments. **, p <0.01. (F) Lifespan of adult male wild-type ( n = 104), parkin P21 ( n = 102) and PGAM5 NP0568 ; parkin P21 ( n = 91) flies. PGAM5 NP0568 ; parkin P21 vs. parkin P21 , p = 0.191; wild-type vs. parkin P21 , p <0.001 by log rank test. (G) Lifespan of adult male parkin P21 ( n = 153) and parkin P21 ; dPGAM5 Tg flies ( n = 155) flies. parkin P21 vs. parkin P21 ; dPGAM5 Tg , p <0.001 by log rank test. (H–K) TEM analysis of the indirect flight muscle and mitochondrial morphology in tissue from flies of the indicated genotypes. The long tubular mitochondrial phenotype seen in parkin P21 flies can be rescued by dPGAM5 inactivation (H and I). However, the mitochondrial matrix still appears degenerated (insets in J and K). Scale bars = 1 µm in H and I and 200 nm in J and K. (L) Quantification of the percentage of mitochondrial size distribution in the indirect muscle tissue from wild-type ( n = 136 from 5 adult flies), parkin P21 ( n = 89 from 5) and parkin P21 ; PGAM5 NP0568 flies ( n = 84 from 5) as shown in (H–K). The length of the mitochondria in the direction of the myofibrils was measured. Data are shown as means ± SE (* p <0.05, ** p <0.01 vs. wild-type ; # p <0.05 vs. parkin P21 ; PGAM5 NP0568 ). The genotypes are: +/Y ( wild-type ), +/Y; parkin P21 /parkin P21 ( parkin P21 ), PGAM5 NP0568 /Y; parkin P21 /parkin P21 ( parkin P21 ; PGAM5 NP0568 ).

Article Snippet: Rabbit anti-human PGAM5 polyclonal antibody was raised against recombinant GST-tagged PGAM5 domain (89–289 aa) produced in the E. coli strain BL21(DE3)pLysS (Novagen), and was affinity-purified against the antigen.

Techniques: Mutagenesis, Activity Assay

Journal: PLoS Genetics

Article Title: The Loss of PGAM5 Suppresses the Mitochondrial Degeneration Caused by Inactivation of PINK1 in Drosophila

doi: 10.1371/journal.pgen.1001229

Figure Lengend Snippet: (A) PGAM5 has a role in mitochondrial activities independently of Parkin downstream of PINK1. (B) PGAM5 negatively regulates Parkin downstream of PINK1.

Article Snippet: Rabbit anti-human PGAM5 polyclonal antibody was raised against recombinant GST-tagged PGAM5 domain (89–289 aa) produced in the E. coli strain BL21(DE3)pLysS (Novagen), and was affinity-purified against the antigen.

Techniques: