human pkm1 cdna  (Addgene inc)

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A , functional enrichment analysis with GO Biological Processes was performed using GOrilla with the differential genes upregulated and downregulated in pulmonary fibroblasts cluster. B , heatmaps are showing the upregulated and downregulated genes of integrin and glycolysis in pulmonary fibroblasts cluster. C , protein expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung homogenates was assessed by immunohistochemistry and quantified by the average optical density value (AOD), n = 3 per group. D , protein expression of integrin β3 and PKM2 in lung homogenates was determined by western blot analysis. Relative densitometry of the protein bands of integrin β3 and PKM2 over β-actin is displayed in bar graphs, n = 3 in sham group and n = 9 in MV group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: After incubation with 10% BSA for 1 h, the sections were incubated with primary antibodies against integrin β3 (SJ1909/NBP2-67416, Novus, USA), PKM2 (AF7244, R&D, USA), fibronectin and collagen-I (72026S, CST, USA) at a dilution of 1:100 at 4 °C overnight.

Techniques: Functional Assay, Expressing, Immunohistochemistry, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A and B, lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker fibronectin (FN) (Alexa Fluor 568, red), integrin β3(β3) (fluorescein isothiocyanate, green) and PKM2 (fluorescein isothiocyanate, green). 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue). Original magnification × 400. Scale bars correspond to 20 µm. C , flow cytometry was used to detect the expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung cells and in fibronectin (FN) positive fibroblasts, n = 6 per group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: After incubation with 10% BSA for 1 h, the sections were incubated with primary antibodies against integrin β3 (SJ1909/NBP2-67416, Novus, USA), PKM2 (AF7244, R&D, USA), fibronectin and collagen-I (72026S, CST, USA) at a dilution of 1:100 at 4 °C overnight.

Techniques: Staining, Labeling, Marker, Flow Cytometry, Expressing

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: Integrin β3 inhibition ameliorates PKM2-mediated aerobic glycolysis and MV-induced pulmonary fibrosis. A and B , both wild-type and integrin β3-deficient (β3 +/- ) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A , lung fibrosis was quantified by Masson's trichrome staining, original magnification × 200, scale bars correspond to 100μm, n = 6, 5, 6, 6, respectively in each group. B , integrin β3, pyruvate kinase M2 (PKM2), lactic dehydrogenase (LDHA), collagen I α1 (COL1A1) and α smooth muscle actin (α-SMA) expression was quantified by western blot analysis, n = 3 per group. C and D , mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4 mg/kg once a day for 3 consecutive days before being subjected to MV for 2 h. Lung tissues were harvested 7 days after intubation. C , lung fibrosis was quantified by Masson's trichrome staining, original magnification × 200, scale bars correspond to 100 µm, n = 6 per group. D , integrin β3, PKM2, LDHA and COL1A1 expression was quantified by western blot analysis, n = 3 per group. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: After incubation with 10% BSA for 1 h, the sections were incubated with primary antibodies against integrin β3 (SJ1909/NBP2-67416, Novus, USA), PKM2 (AF7244, R&D, USA), fibronectin and collagen-I (72026S, CST, USA) at a dilution of 1:100 at 4 °C overnight.

Techniques: Inhibition, Staining, Expressing, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: Integrin β3 inhibition ameliorates PKM2-mediated aerobic glycolysis and MV-induced pulmonary fibrosis . A-D , both wild-type and integrin β3-deficient (β3 +/- ) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A and B , lung tissues were stained with fluorophorelabeled antibodies against fibroblast marker fibronectin (FN) (red), pyruvate kinase M2 (PKM2) (green) and collagen I (green), 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. C and D , lactate (Lac) and type I procollagen carboxyterminal peptide (PICP) levels in bronchoalveolar lavage fluid (BALF) were tested, n = 6, 3, 6, 3, respectively in each group. E-H , mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4mg/kg once a day for 3 consecutive days before being subjected to MV for 2h. Lung tissues were harvested 7days after intubation. E and F , Lac and PICP were tested in BALF, n = 6, 4, 6, 6, respectively in each group. G and H , lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker FN (red), PKM2 (green) and collagen I (green), DAPI stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: After incubation with 10% BSA for 1 h, the sections were incubated with primary antibodies against integrin β3 (SJ1909/NBP2-67416, Novus, USA), PKM2 (AF7244, R&D, USA), fibronectin and collagen-I (72026S, CST, USA) at a dilution of 1:100 at 4 °C overnight.

Techniques: Inhibition, Staining, Marker, Labeling

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A , functional enrichment analysis with GO Biological Processes was performed using GOrilla with the differential genes upregulated and downregulated in pulmonary fibroblasts cluster. B , heatmaps are showing the upregulated and downregulated genes of integrin and glycolysis in pulmonary fibroblasts cluster. C , protein expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung homogenates was assessed by immunohistochemistry and quantified by the average optical density value (AOD), n = 3 per group. D , protein expression of integrin β3 and PKM2 in lung homogenates was determined by western blot analysis. Relative densitometry of the protein bands of integrin β3 and PKM2 over β-actin is displayed in bar graphs, n = 3 in sham group and n = 9 in MV group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Functional Assay, Expressing, Immunohistochemistry, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: MV leads to upregulation of integrin β3 and PKM2 in pulmonary fibroblasts. Mice lung tissue were acquired at Day 7 after 2 h of MV. A and B, lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker fibronectin (FN) (Alexa Fluor 568, red), integrin β3(β3) (fluorescein isothiocyanate, green) and PKM2 (fluorescein isothiocyanate, green). 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue). Original magnification × 400. Scale bars correspond to 20 µm. C , flow cytometry was used to detect the expression of integrin β3 and pyruvate kinase M2 (PKM2) in lung cells and in fibronectin (FN) positive fibroblasts, n = 6 per group. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, *** p < 0.001(t test).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Staining, Labeling, Marker, Flow Cytometry, Expressing

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: Integrin β3 inhibition ameliorates PKM2-mediated aerobic glycolysis and MV-induced pulmonary fibrosis. A and B , both wild-type and integrin β3-deficient (β3 +/- ) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A , lung fibrosis was quantified by Masson's trichrome staining, original magnification × 200, scale bars correspond to 100μm, n = 6, 5, 6, 6, respectively in each group. B , integrin β3, pyruvate kinase M2 (PKM2), lactic dehydrogenase (LDHA), collagen I α1 (COL1A1) and α smooth muscle actin (α-SMA) expression was quantified by western blot analysis, n = 3 per group. C and D , mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4 mg/kg once a day for 3 consecutive days before being subjected to MV for 2 h. Lung tissues were harvested 7 days after intubation. C , lung fibrosis was quantified by Masson's trichrome staining, original magnification × 200, scale bars correspond to 100 µm, n = 6 per group. D , integrin β3, PKM2, LDHA and COL1A1 expression was quantified by western blot analysis, n = 3 per group. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Inhibition, Staining, Expressing, Western Blot

Journal: Theranostics

Article Title: Integrin β3-PKM2 pathway-mediated aerobic glycolysis contributes to mechanical ventilation-induced pulmonary fibrosis

doi: 10.7150/thno.72328

Figure Lengend Snippet: Integrin β3 inhibition ameliorates PKM2-mediated aerobic glycolysis and MV-induced pulmonary fibrosis . A-D , both wild-type and integrin β3-deficient (β3 +/- ) mice were subjected to mechanical ventilation (MV) for 2 h. Lung tissues were harvested at day 7. A and B , lung tissues were stained with fluorophorelabeled antibodies against fibroblast marker fibronectin (FN) (red), pyruvate kinase M2 (PKM2) (green) and collagen I (green), 4',6-diamidino-2- phenylindole (DAPI) stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. C and D , lactate (Lac) and type I procollagen carboxyterminal peptide (PICP) levels in bronchoalveolar lavage fluid (BALF) were tested, n = 6, 3, 6, 3, respectively in each group. E-H , mice were treated intraperitoneally with integrin β3 inhibitor cilengitide 4mg/kg once a day for 3 consecutive days before being subjected to MV for 2h. Lung tissues were harvested 7days after intubation. E and F , Lac and PICP were tested in BALF, n = 6, 4, 6, 6, respectively in each group. G and H , lung tissues were stained with fluorophore-labeled antibodies against fibroblast marker FN (red), PKM2 (green) and collagen I (green), DAPI stain was used to detect nuclei (blue), original magnification × 400, scale bars correspond to 20 µm. Data are expressed as means ± SEM, *p < 0.05, **p < 0.01, *** p < 0.001 (ANOVA).

Article Snippet: Mouse monoclonal integrin β3 (SJ1909/NBP2-67416, Novus, USA) and PKM2 antibody (AF7244, R&D, USA) were added on the sections as primary antibodies.

Techniques: Inhibition, Staining, Marker, Labeling

Journal: Biomolecules

Article Title: PTEN Protein Phosphatase Activity Is Not Required for Tumour Suppression in the Mouse Prostate.

doi: 10.3390/biom12101511

Figure Lengend Snippet: Figure 5. Representative immunohistochemistry showing the abundance of Keratin 19 (KRT19) and Pyruvate Kinase M2 (PKM2) in sections of prostate tissues from wild-type (W/W) and mice conditionally deleted for Pten in the prostate (F/F) at 6 weeks of age. W/W = Pten+/+; F/F = Ptenflox/flox. Scale bars: 50 µm.

Article Snippet: Antibodies and Immunohistochemistry: The antibodies used for Immunohistochemistry (IHC) were: Phospho-S473 AKT Antibody #9271 from Cell Signaling Technology (also used for immunoblotting); PTEN antibody Clone 138G6 #9559 from Cell Signalling Technology; Androgen Receptor (AR) from DAKO; PKM2 antibody from R&D systems (AF7244); RAC1-GTP mAb from NewEastBio #26903.

Techniques: Immunohistochemistry

Journal: Cell metabolism

Article Title: Warburg-like metabolic transformation underlies neuronal degeneration in sporadic Alzheimer's disease.

doi: 10.1016/j.cmet.2022.07.014

Figure Lengend Snippet: Figure 5. Nuclear PKM2 activity alters the neuronal epigenetic landscape (A) Schematic: phosphorylated PKM2 translocates to the nucleus to interact with transcription factors to regulate gene expression. (B and C) Immunostaining (B) and quantification (C) of p-PKM2(Ser37) (control, n = 8; AD, n = 5). Scale bars, 10 mm. (D and E) Immunostaining (D) and quantification (E) of phosphorylated histone 3 (T11) of MAP2-positive neurons (control, n = 6; AD, n = 5). Scale bars, 10 mm. (F) ATAC-seq profiles around transcriptional start sites of genes regulated by HIF1a, STAT3, and b-catenin (CTNNB1), based on ReMap2020 (control, n = 11; AD, n = 9). (G) HOMER motif enrichment analysis of AD differentially open peaks for HIF1a and STAT3, as previously published (Mertens et al., 2021). (H and I) Differential expression (H) and GO term enrichment (I) of significant genes regulated by HIF1a or STAT3. (B–H) Violin plots: median and quartiles. Significance: unpaired t test, *p < 0.05, **p < 0.01.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti Beta-III-tubulin BioLegend Cat#MMS-435P; RRID:AB_2313773 Rabbit anti Beta-III-tubulin BioLegend Cat#802001; RRID:AB_2564645 Mouse anti NeuN EMD Millipore Cat#MAB377; RRID:AB_2298772 Rabbit anti NeuN Cell Signaling Cat#24307T; RRID:AB_2651140 PE-conjugated anti PSA-NCAM Miltenyi Biotec Cat#130-117-394; RRID:AB_2727931 Rabbit anti Synapsin Merck Cat#574778; RRID:AB_565174 Mouse anti PSD-95 (clone K28/42) NeuroMab Cat#75028; RRID:AB_2292909 Chicken anti Map2 Abcam Cat#ab5392; RRID:AB_2138153 Rabbit PKM2 (phospho-Ser37) Sabbiotech Cat#11456 Mouse PKM2 R&D systems Cat#MAB72441 Rabbit Histone 3 (phospho T11) Abcam Cat#ab5168, RRID:AB_304759 Rabbit cleaved caspase 3 (asp175)(5A1E) Cell Signaling Cat#9664; RRID:AB_2070042 Mouse PDH-E1a Santa Cruz Cat#sc-377092; RRID:AB_2716767 Mouse PKM2 ThermoFisher Scientific Cat#TA190266 Alexa Fluor 488-conjugated Donkey Anti Rabbit IgG Thermo Fisher Scientific Cat#A-21206; RRID:AB_2535792 Alexa Fluor 647-conjugated Donkey Anti Rabbit IgG Thermo Fisher Scientific Cat#A-31573; RRID:AB_2536183 Alexa Fluor 488-conjugated Donkey Anti Mouse IgG Thermo Fisher Scientific Cat#A-21202; RRID:AB_141607 Alexa Fluor 647-conjugated Donkey Anti Mouse IgG Thermo Fisher Scientific Cat#A-31571; RRID:AB_162542 Alexa Fluor 647-conjugated Donkey Anti Chicken IgY Millipore Cat#AP194SA6; RRID:AB_2650475 Cy3-conjugated Donkey Anti-Mouse IgG Jackson ImmunoRes.

Techniques: Activity Assay, Gene Expression, Immunostaining, Control, Quantitative Proteomics

Journal: Cell metabolism

Article Title: Warburg-like metabolic transformation underlies neuronal degeneration in sporadic Alzheimer's disease.

doi: 10.1016/j.cmet.2022.07.014

Figure Lengend Snippet: Figure 6. A metabolic shift induces AD-like apoptotic competency in human neurons (A and B) Immunostaining (A) and quantification (B) of cleaved caspase-3 over DAPI of bIII-tub-positive control and AD iNs (control, n = 9; AD, n = 7). Scale bars, 50 mm. Green arrows point out Casp3-positive neurons. (C) Schematic: induction of neuronal apoptosis by ABT-737 treatment. (D) Cell death assessed by cleaved caspase 3/bIII-tub-positive cells of control (green) and AD (teal) iNs in response to 0–1 mM ABT-737. (E and F) Immunostaining (E) and quantification (F) of cleaved caspase-3 in bIII-tub neurons after Bcl2 inhibition in control (n = 10) and AD (n = 8) iNs. Scale bars, 50 mm. (G) Pearson correlation analysis of cleaved caspase-3 immunostaining and glycolytic metabolites (UHPLC-MS). (H and I) Control iNs treated with CoDo for 2 days (H) showed increased lactate secretion (vehicle, n = 5; CoDo, n = 5) (I). (J) Quantification of p-PKM2 FI in the nucleus/cytoplasm comparing vehicle-treated (n = 6) and CoDo-treated control iNs (n = 6). (K) Immunostainings of bIII-tub and EGFP fluorescence of EGFP::PKM2 transduced iNs. Dotted lines show cytoplasmic ROI. Scale bars, 100 and 25 mm. (L) Longitudinal EGFP::PKM2 localization in vehicle-treated (n = 3) or CoCl2-treated (n = 4) control iNs. (M and N) Immunostaining (M) and quantification (N) of cleaved caspase-3 positive cells/DAPI before and after ABT-737 (vehicle and CoDo, n = 6; one-way ANOVA, DF: 23, F = 21.34, p < 0.0001, Dunnett’s multiple comparison). Scale bars, 50 mm. Dots represent individual donors throughout the figure. Bars, mean; error bars, SD; significance, unpaired t test, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti Beta-III-tubulin BioLegend Cat#MMS-435P; RRID:AB_2313773 Rabbit anti Beta-III-tubulin BioLegend Cat#802001; RRID:AB_2564645 Mouse anti NeuN EMD Millipore Cat#MAB377; RRID:AB_2298772 Rabbit anti NeuN Cell Signaling Cat#24307T; RRID:AB_2651140 PE-conjugated anti PSA-NCAM Miltenyi Biotec Cat#130-117-394; RRID:AB_2727931 Rabbit anti Synapsin Merck Cat#574778; RRID:AB_565174 Mouse anti PSD-95 (clone K28/42) NeuroMab Cat#75028; RRID:AB_2292909 Chicken anti Map2 Abcam Cat#ab5392; RRID:AB_2138153 Rabbit PKM2 (phospho-Ser37) Sabbiotech Cat#11456 Mouse PKM2 R&D systems Cat#MAB72441 Rabbit Histone 3 (phospho T11) Abcam Cat#ab5168, RRID:AB_304759 Rabbit cleaved caspase 3 (asp175)(5A1E) Cell Signaling Cat#9664; RRID:AB_2070042 Mouse PDH-E1a Santa Cruz Cat#sc-377092; RRID:AB_2716767 Mouse PKM2 ThermoFisher Scientific Cat#TA190266 Alexa Fluor 488-conjugated Donkey Anti Rabbit IgG Thermo Fisher Scientific Cat#A-21206; RRID:AB_2535792 Alexa Fluor 647-conjugated Donkey Anti Rabbit IgG Thermo Fisher Scientific Cat#A-31573; RRID:AB_2536183 Alexa Fluor 488-conjugated Donkey Anti Mouse IgG Thermo Fisher Scientific Cat#A-21202; RRID:AB_141607 Alexa Fluor 647-conjugated Donkey Anti Mouse IgG Thermo Fisher Scientific Cat#A-31571; RRID:AB_162542 Alexa Fluor 647-conjugated Donkey Anti Chicken IgY Millipore Cat#AP194SA6; RRID:AB_2650475 Cy3-conjugated Donkey Anti-Mouse IgG Jackson ImmunoRes.

Techniques: Immunostaining, Positive Control, Control, Inhibition, Comparison

Journal: Cell metabolism

Article Title: Warburg-like metabolic transformation underlies neuronal degeneration in sporadic Alzheimer's disease.

doi: 10.1016/j.cmet.2022.07.014

Figure Lengend Snippet: Figure 7. PKM2 inhibition ameliorates PKM2-induced apoptotic competency (A) Schematic: shikonin treatment prevents PKM2 nuclear translocation and increases metabolic enzymatic activity. (B) Longitudinal EGFP::PKM2 localization in vehicle-treated (n = 3), CoCl2-treated (n = 4), and CoCl2+shikonin-treated (n = 4) control iNs.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti Beta-III-tubulin BioLegend Cat#MMS-435P; RRID:AB_2313773 Rabbit anti Beta-III-tubulin BioLegend Cat#802001; RRID:AB_2564645 Mouse anti NeuN EMD Millipore Cat#MAB377; RRID:AB_2298772 Rabbit anti NeuN Cell Signaling Cat#24307T; RRID:AB_2651140 PE-conjugated anti PSA-NCAM Miltenyi Biotec Cat#130-117-394; RRID:AB_2727931 Rabbit anti Synapsin Merck Cat#574778; RRID:AB_565174 Mouse anti PSD-95 (clone K28/42) NeuroMab Cat#75028; RRID:AB_2292909 Chicken anti Map2 Abcam Cat#ab5392; RRID:AB_2138153 Rabbit PKM2 (phospho-Ser37) Sabbiotech Cat#11456 Mouse PKM2 R&D systems Cat#MAB72441 Rabbit Histone 3 (phospho T11) Abcam Cat#ab5168, RRID:AB_304759 Rabbit cleaved caspase 3 (asp175)(5A1E) Cell Signaling Cat#9664; RRID:AB_2070042 Mouse PDH-E1a Santa Cruz Cat#sc-377092; RRID:AB_2716767 Mouse PKM2 ThermoFisher Scientific Cat#TA190266 Alexa Fluor 488-conjugated Donkey Anti Rabbit IgG Thermo Fisher Scientific Cat#A-21206; RRID:AB_2535792 Alexa Fluor 647-conjugated Donkey Anti Rabbit IgG Thermo Fisher Scientific Cat#A-31573; RRID:AB_2536183 Alexa Fluor 488-conjugated Donkey Anti Mouse IgG Thermo Fisher Scientific Cat#A-21202; RRID:AB_141607 Alexa Fluor 647-conjugated Donkey Anti Mouse IgG Thermo Fisher Scientific Cat#A-31571; RRID:AB_162542 Alexa Fluor 647-conjugated Donkey Anti Chicken IgY Millipore Cat#AP194SA6; RRID:AB_2650475 Cy3-conjugated Donkey Anti-Mouse IgG Jackson ImmunoRes.

Techniques: Inhibition, Translocation Assay, Activity Assay, Control