Anti-DRP1 (phospho S637) antibody

• WB

Supplier Data

Western blot - Anti-DRP1 (phospho S637) antibody (AB193216)

All lanes:

Western blot - Anti-DRP1 (phospho S637) antibody (ab193216)

Lane 1:

293 whole cell lysates with phospho-blocking peptide (P-Peptide)

Lane 2:

293 whole cell lysates with non-phospho-blocking peptide (N-Peptide)

Lane 3:

293 whole cell lysates

Predicted band size: 82 kDa

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• WB

Supplier Data

Western blot - Anti-DRP1 (phospho S637) antibody (AB193216)

All lanes:

Western blot - Anti-DRP1 (phospho S637) antibody (ab193216) at 1/500 dilution

Lane 1:

Hela whole cell lysate at 40 µg

Lane 2:

Hela whole cell lysate treated with 100nM Calyculin A for 30min at 40 µg

Lane 3:

Hela whole cell lysate treated with 10 µM Forskolin for 30min at 40 µg

Secondary

All lanes:

All lanes : Goat Anti-Rabbit IgG at 1/20000 dilution

Predicted band size: 82 kDa

Observed band size: 82 kDa

true

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-DRP1 (phospho S637) antibody (AB193216)

All lanes:

Western blot - Anti-DRP1 (phospho S637) antibody (ab193216) at 1/500 dilution

Lane 1:

Jurkat (Human T cell leukemia cell line from peripheral blood) cell extract

Lane 2:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell extract

Lane 3:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) cell extract with antigen-specific peptide

Predicted band size: 82 kDa

false

• WB

CiteAb

Western blot - Anti-DRP1 (phospho S637) antibody (AB193216)

Western Blotting using Anti-DRP1 (phospho S637) antibody, ab193216. Publication image from Yu, B. et al., 2020, Nat Commun, 32439975. Legend direct from paper.

Mitochondrial dynamics and function change upon PGAM5 deletion.a Mitochondrial morphology outlined by Tom20 antibodies in control and PGAM5−/− ARPE-19 cells. Scale bar = 20 µm. n = 5. b Western blots showing upregulation of phosphor-Drp1(S637) but not total Drp1 in PGAM5−/− ARPE-19 cells.α-Tubulin was used as loading control. n = 4. c Western blots showing PGAM5 cleavage and Drp1(S637) dephosphorylation in ARPE-19 cells by CCCP treatment. n = 3. d Co-immunoprecipitation experiment using Axin1 antibody, showing that Axin1 interacts with both Drp1 and cleaved PGAM5 in ARPE-19 cells. n = 3. e Western blots showing upregulation of mitochondrial proteins (Tom20, CYTC, CYPD) and downregulation of PGC1α in PGAM5−/− ARPE-19 cells.α-Tubulin was used as loading control. n = 4. f Increased mitochondrial DNA in PGAM5−/− ARPE-19 cells. n = 5. *p = 0.0111, two-tailed unpaired t-tests; error bars, mean ± s.e.m. g Increased mitochondrial protein Cypd in the RPE/choroid of Pgam5−/− mice.α-Tubulin was used as loading control. n = 3. h Decreased mitochondrial turnover in PGAM5−/− ARPE-19 by MitoTimer transfection and labeling. Scale Bar equals to 20 µm. n = 3. i Mitochondrial membrane potential change as labeled by JC-1 in WT and PGAM5−/− ARPE-19 cells. Scale bar = 20 µm. n = 5. j ATP level change as measured in short-term (1 week) and long-term (8 weeks) culture of WT and PGAM5−/− ARPE-19 cells. n = 3, ****p < 0.0001, two-way ANOVA Tukey’s multiple comparisons test; error bars, mean ± s.e.m. k ROS change as measured in short-term (1 week) and long-term (8 weeks) culture of WT and PGAM5−/− ARPE-19 cells. n = 3, *p < 0.05; ****p < 0.0001, two-way ANOVA Tukey’s multiple comparisons test. n.s. represents no significance. Error bars, mean ± s.e.m.; for assays in the figure, n represents the number of biologically independent experiments. Images were captured under same settings, and representative images were shown. Source data are available as a Source Data file.

false

• WB

CiteAb

Western blot - Anti-DRP1 (phospho S637) antibody (AB193216)

Western Blotting using Anti-DRP1 (phospho S637) antibody, ab193216. Publication image from Yu, B. et al., 2020, Nat Commun, 32439975. Legend direct from paper.

Drp1-K38A overexpression phenocopies PGAM5−/− mitochondrial phenotype and cell senescence.a Tom20 immunostaining in WT and PGAM5−/− ARPE-19 cells infected with adenovirus expressing Ad-LacZ or Drp1-K38A mutant, showing that Ad-Drp1-K38A mutant mimics PGAM5−/− hyperfusion phenotype. Scale bar = 20 µm. n = 4. b Quantification of mitochondrial branches in a using ImageJ. n = 4, *p < 0.05, **p < 0.005, analyzed by two-way ANOVA Tukey’s multiple comparisons test. Error bars, mean ± s.e.m. c ATP production in WT ARPE-19 cells overexpressing Ad-LacZ or Ad-Drp1-K38A. n = 10 biologically independent samples, ****p < 0.0001, two-tailed unpaired t-tests, error bars, mean ± s.d. d ATP production in PGAM5 KO cells overexpressing Ad-LacZ or Ad-Drp1-K38A. n = 10 biologically independent samples, ****p < 0.0001, two-tailed unpaired t-tests, error bars, mean ± s.d. e Expression of DRP1, phosphor-S6 and S6 in Ad-LacZ or Ad-Drp1-K38A overexpressing HRPE cells at 7 days after infection. Samples were collected at 48 h after the last medium change for western blot. β-Actin was used as loading control. n = 3. f SA-gal staining of HRPE cells 5 weeks after Ad-GFP or Ad-Drp1-K38A infection (MOI = 100). Scale bar = 500 µm. Boxed region represents the magnified picture in the figure. n = 3. g Expression of Drp1, Lamin B1, MMP3 and P16Ink4a protein at 8 weeks after infection. Samples were collected at 48 h after the last medium change for western blot.α-Tubulin was used as loading control. n = 4. h Quantification of bands in g. n = 4, **p = 0.0018 (Lamin B1), **p = 0.0026 (MMP3), ***p = 0.0004, two-tailed unpaired t-tests, error bars, mean ± s.d. i mRNA level of IL6, MMP3 and TNFα were measured by qRT-PCR at 8 weeks after infection as shown in f. Samples were collected at 48 h after the last medium change. n = 3, *p = 0.0338, **p = 0.0057, ***p = 0.0001, two-tailed unpaired t-tests, error bars, mean ± s.e.m. For assays in the figure, n represents the number of biologically independent experiments unless otherwise specified. Images were captured under same settings, and representative images were shown. Source data are available as a Source Data file.

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