ps 1  (Danaher Inc)


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    Danaher Inc ps 1
    Experimental procedure and targeted molecular signaling. A Experimental protocol; B C3G-mediated amyloidogenic pathway and tau protein phosphorylation; C C3G enhanced autophagy in APPswe mice; D neuroprotective effects of C3G on neuronal apoptosis; and E C3G improved the synaptic function. APP, amyloid precursor protein; BACE1, β-secretase; <t>PS-1,</t> presenilin-1; ADAM10, α-secretase; Aβ, amyloid beta; IL, interleukin; TNF-α, tumor necrosis factor-α; GSH-Px, glutathione peroxidase; CAT, catalase; NFTs, neurofibrillary tangles; C3G, cyaninidin-3-O-glucoside; Bax, Bcl-2 associated X, apoptosis regulator; Bcl2, B-cell lymphoma 2; MAPKs, mitogen-activated protein kinases; GSK3β, glycogen synthase kinase-3 beta; AMPK, AMP-activated protein kinase; SIRT1, sirtuin 1; PI3K/Akt, phosphoinositide 3-kinase/protein kinase B; SYP, synaptophysin; PSD95, postsynaptic density protein-95; LC3B-II, light chain 3BII; LAMP-1, lysosomal-associated membrane protein 1; TFEB, transcription factor EB; SQSTM, sequestosome 1.
    Ps 1, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ps 1/product/Danaher Inc
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    1) Product Images from "Cyanidin-3- O -glucoside protects the brain and improves cognitive function in APPswe/PS1ΔE9 transgenic mice model"

    Article Title: Cyanidin-3- O -glucoside protects the brain and improves cognitive function in APPswe/PS1ΔE9 transgenic mice model

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-023-02950-3

    Experimental procedure and targeted molecular signaling. A Experimental protocol; B C3G-mediated amyloidogenic pathway and tau protein phosphorylation; C C3G enhanced autophagy in APPswe mice; D neuroprotective effects of C3G on neuronal apoptosis; and E C3G improved the synaptic function. APP, amyloid precursor protein; BACE1, β-secretase; PS-1, presenilin-1; ADAM10, α-secretase; Aβ, amyloid beta; IL, interleukin; TNF-α, tumor necrosis factor-α; GSH-Px, glutathione peroxidase; CAT, catalase; NFTs, neurofibrillary tangles; C3G, cyaninidin-3-O-glucoside; Bax, Bcl-2 associated X, apoptosis regulator; Bcl2, B-cell lymphoma 2; MAPKs, mitogen-activated protein kinases; GSK3β, glycogen synthase kinase-3 beta; AMPK, AMP-activated protein kinase; SIRT1, sirtuin 1; PI3K/Akt, phosphoinositide 3-kinase/protein kinase B; SYP, synaptophysin; PSD95, postsynaptic density protein-95; LC3B-II, light chain 3BII; LAMP-1, lysosomal-associated membrane protein 1; TFEB, transcription factor EB; SQSTM, sequestosome 1.
    Figure Legend Snippet: Experimental procedure and targeted molecular signaling. A Experimental protocol; B C3G-mediated amyloidogenic pathway and tau protein phosphorylation; C C3G enhanced autophagy in APPswe mice; D neuroprotective effects of C3G on neuronal apoptosis; and E C3G improved the synaptic function. APP, amyloid precursor protein; BACE1, β-secretase; PS-1, presenilin-1; ADAM10, α-secretase; Aβ, amyloid beta; IL, interleukin; TNF-α, tumor necrosis factor-α; GSH-Px, glutathione peroxidase; CAT, catalase; NFTs, neurofibrillary tangles; C3G, cyaninidin-3-O-glucoside; Bax, Bcl-2 associated X, apoptosis regulator; Bcl2, B-cell lymphoma 2; MAPKs, mitogen-activated protein kinases; GSK3β, glycogen synthase kinase-3 beta; AMPK, AMP-activated protein kinase; SIRT1, sirtuin 1; PI3K/Akt, phosphoinositide 3-kinase/protein kinase B; SYP, synaptophysin; PSD95, postsynaptic density protein-95; LC3B-II, light chain 3BII; LAMP-1, lysosomal-associated membrane protein 1; TFEB, transcription factor EB; SQSTM, sequestosome 1.

    Techniques Used: Membrane

    antibody against pser129 ps α syn  (Danaher Inc)


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    Danaher Inc antibody against pser129 ps α syn
    Antibody Against Pser129 Ps α Syn, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody against pser129 ps α syn/product/Danaher Inc
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    Structured Review

    Abcam ps pkc substrate
    ( A ) Effects of various kinase inhibitors on the β CBP promoter via dual luciferase reporter assays. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells overexpressing ATF2 and incubated with the <t>PKC</t> inhibitor (20 μM), ERK inhibitor (30 μM), p38 inhibitor (30 μM), or JNK inhibitor (30 μM). DMSO was used as a control. P < 0.05, n = 3. ( B ) Relative levels of β CBP and ATF2 in the Pron of the fifth-instar gregarious locust nymphs injected with various inhibitors. Inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. * P < 0.05, n = 5 or 6. ( C ) Verification of ATF2 phosphorylation at serine stimulated by PKC. PKC inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. PBS was used as a control. ATF2 protein was enriched from the Pron of the fifth-instar gregarious locust nymphs using anti-ATF2 antibody-bound CNBr-activated Sepharose 4B. The phosphorylation of ATF2 was detected using anti–phospho-(Ser) PKC substrate (pS-PKC substrate) <t>polyclonal</t> <t>antibodies.</t> * P < 0.05 and ** P < 0.01, n = 4. ( D ) Expression is driven by the promoter of β CBP in the presence of various constructs. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells with ATF2 (WT), ATF2 S45A, ATF2 S327A, ATF2 S406A, ATF2 S518A, and ATF2 S668A, respectively. * P < 0.05, n = 3. Fluc/Rluc represents the fold of firefly to Renilla luciferase activity. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).
    Ps Pkc Substrate, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ps pkc substrate - by Bioz Stars, 2023-11
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    Images

    1) Product Images from "Spatially differential regulation of ATF2 phosphorylation contributes to warning coloration of gregarious locusts"

    Article Title: Spatially differential regulation of ATF2 phosphorylation contributes to warning coloration of gregarious locusts

    Journal: Science Advances

    doi: 10.1126/sciadv.adi5168

    ( A ) Effects of various kinase inhibitors on the β CBP promoter via dual luciferase reporter assays. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells overexpressing ATF2 and incubated with the PKC inhibitor (20 μM), ERK inhibitor (30 μM), p38 inhibitor (30 μM), or JNK inhibitor (30 μM). DMSO was used as a control. P < 0.05, n = 3. ( B ) Relative levels of β CBP and ATF2 in the Pron of the fifth-instar gregarious locust nymphs injected with various inhibitors. Inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. * P < 0.05, n = 5 or 6. ( C ) Verification of ATF2 phosphorylation at serine stimulated by PKC. PKC inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. PBS was used as a control. ATF2 protein was enriched from the Pron of the fifth-instar gregarious locust nymphs using anti-ATF2 antibody-bound CNBr-activated Sepharose 4B. The phosphorylation of ATF2 was detected using anti–phospho-(Ser) PKC substrate (pS-PKC substrate) polyclonal antibodies. * P < 0.05 and ** P < 0.01, n = 4. ( D ) Expression is driven by the promoter of β CBP in the presence of various constructs. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells with ATF2 (WT), ATF2 S45A, ATF2 S327A, ATF2 S406A, ATF2 S518A, and ATF2 S668A, respectively. * P < 0.05, n = 3. Fluc/Rluc represents the fold of firefly to Renilla luciferase activity. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).
    Figure Legend Snippet: ( A ) Effects of various kinase inhibitors on the β CBP promoter via dual luciferase reporter assays. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells overexpressing ATF2 and incubated with the PKC inhibitor (20 μM), ERK inhibitor (30 μM), p38 inhibitor (30 μM), or JNK inhibitor (30 μM). DMSO was used as a control. P < 0.05, n = 3. ( B ) Relative levels of β CBP and ATF2 in the Pron of the fifth-instar gregarious locust nymphs injected with various inhibitors. Inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. * P < 0.05, n = 5 or 6. ( C ) Verification of ATF2 phosphorylation at serine stimulated by PKC. PKC inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. PBS was used as a control. ATF2 protein was enriched from the Pron of the fifth-instar gregarious locust nymphs using anti-ATF2 antibody-bound CNBr-activated Sepharose 4B. The phosphorylation of ATF2 was detected using anti–phospho-(Ser) PKC substrate (pS-PKC substrate) polyclonal antibodies. * P < 0.05 and ** P < 0.01, n = 4. ( D ) Expression is driven by the promoter of β CBP in the presence of various constructs. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells with ATF2 (WT), ATF2 S45A, ATF2 S327A, ATF2 S406A, ATF2 S518A, and ATF2 S668A, respectively. * P < 0.05, n = 3. Fluc/Rluc represents the fold of firefly to Renilla luciferase activity. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).

    Techniques Used: Luciferase, Incubation, Injection, Expressing, Construct, Activity Assay, Two Tailed Test

    ( A ) β CBP and ATF2 mRNA levels after PKC α knockdown in Pron. PKC α knockdown (3 μg per nymph administered in the day 2 of the second-instar gregarious locust nymphs, four times over 48-hours intervals). dsGFP was used as a control. ** P < 0.01, n = 4. ( B ) ATF2 phosphorylation levels in Pron after PKC isoform knockdown. ** P < 0.01, n = 3 or 4. ( C ) Body color phenotypes of gregarious locust nymphs after RNAi exposure. Images were obtained on the fifth-instar nymphs after RNAi exposure. Red box represents locusts with a notable black-brown pattern. Blue box represents locusts with a cryptic green color. Orange box represents locusts with a body color that falls between the notable black-brown pattern and the cryptic green color. Scale bar, 0.5 cm. * P < 0.05 and ** P < 0.01, n = 3. ( D ) Immunohistochemistry of ATF2 and pS-ATF2 in Pros after PKC α knockdown. Green denotes ATF2 or pS-ATF2 staining by anti-ATF2 antibodies or anti–phospho-ATF2 (Ser 327 ) antibodies. Blue denotes DAPI staining for nuclei. Scale bars, 25 μm. ( E ) β-carotene content coupled with βCBP was evaluated in Pron after PKC α knockdown by HPLC. * P < 0.05, n = 5. ( F ) ChIP assays show that PKC α knockdown repressed the relative precipitation of the β CBP promoter region in Pros. pS-ATF2, phospho-ATF2 (Ser 327 ) antibodies; ATF2, ATF2 antibodies; IgG, nonspecific rabbit IgG control. P < 0.05, n = 3 or 4. Pron: The fifth-instar gregarious locust nymphs pronotum. Pros: The fifth-instar gregarious locust nymphs prosternum. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).
    Figure Legend Snippet: ( A ) β CBP and ATF2 mRNA levels after PKC α knockdown in Pron. PKC α knockdown (3 μg per nymph administered in the day 2 of the second-instar gregarious locust nymphs, four times over 48-hours intervals). dsGFP was used as a control. ** P < 0.01, n = 4. ( B ) ATF2 phosphorylation levels in Pron after PKC isoform knockdown. ** P < 0.01, n = 3 or 4. ( C ) Body color phenotypes of gregarious locust nymphs after RNAi exposure. Images were obtained on the fifth-instar nymphs after RNAi exposure. Red box represents locusts with a notable black-brown pattern. Blue box represents locusts with a cryptic green color. Orange box represents locusts with a body color that falls between the notable black-brown pattern and the cryptic green color. Scale bar, 0.5 cm. * P < 0.05 and ** P < 0.01, n = 3. ( D ) Immunohistochemistry of ATF2 and pS-ATF2 in Pros after PKC α knockdown. Green denotes ATF2 or pS-ATF2 staining by anti-ATF2 antibodies or anti–phospho-ATF2 (Ser 327 ) antibodies. Blue denotes DAPI staining for nuclei. Scale bars, 25 μm. ( E ) β-carotene content coupled with βCBP was evaluated in Pron after PKC α knockdown by HPLC. * P < 0.05, n = 5. ( F ) ChIP assays show that PKC α knockdown repressed the relative precipitation of the β CBP promoter region in Pros. pS-ATF2, phospho-ATF2 (Ser 327 ) antibodies; ATF2, ATF2 antibodies; IgG, nonspecific rabbit IgG control. P < 0.05, n = 3 or 4. Pron: The fifth-instar gregarious locust nymphs pronotum. Pros: The fifth-instar gregarious locust nymphs prosternum. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).

    Techniques Used: Immunohistochemistry, Staining, Two Tailed Test


    Structured Review

    Abcam rabbit anti ps
    Rabbit Anti Ps, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    rabbit anti ps - by Bioz Stars, 2023-11
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    anti ps 129 α syn monoclonal antibody ep1536y  (Abcam)

     
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    Abcam anti ps 129 α syn monoclonal antibody ep1536y
    Measurement of <t>pS</t> <t>129</t> <t>-α-syn</t> in human and mouse samples. All measurements were performed using biotinylated <t>EP1536Y</t> for capture and MSD’s Sulfo-tagged anti-human-α-syn antibody for detection. (A) Measurement of pS 129 -α-syn in human CSF, serum, plasma, and saliva. HC, healthy control; PD, Parkinson’s disease; DLB, dementia with Lewy bodies; MSA, multiple system atrophy; LLoD, lower limit of detection; LLoQ, lower limit of quantitation. The p -values were calculated by a one-way ANOVA with post hoc Tukey test separately for the CSF samples and the serum, plasma, and saliva samples. (B) Comparison of pS 129 -α-syn concentrations in brain extracts from wild-type and MSA model mice. The p -values were calculated by Mann–Whitney test. The data are shown as the mean ± SD.
    Anti Ps 129 α Syn Monoclonal Antibody Ep1536y, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti ps 129 α syn monoclonal antibody ep1536y/product/Abcam
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    1) Product Images from "Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples"

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    Journal: ACS Chemical Neuroscience

    doi: 10.1021/acschemneuro.2c00676

    Measurement of pS 129 -α-syn in human and mouse samples. All measurements were performed using biotinylated EP1536Y for capture and MSD’s Sulfo-tagged anti-human-α-syn antibody for detection. (A) Measurement of pS 129 -α-syn in human CSF, serum, plasma, and saliva. HC, healthy control; PD, Parkinson’s disease; DLB, dementia with Lewy bodies; MSA, multiple system atrophy; LLoD, lower limit of detection; LLoQ, lower limit of quantitation. The p -values were calculated by a one-way ANOVA with post hoc Tukey test separately for the CSF samples and the serum, plasma, and saliva samples. (B) Comparison of pS 129 -α-syn concentrations in brain extracts from wild-type and MSA model mice. The p -values were calculated by Mann–Whitney test. The data are shown as the mean ± SD.
    Figure Legend Snippet: Measurement of pS 129 -α-syn in human and mouse samples. All measurements were performed using biotinylated EP1536Y for capture and MSD’s Sulfo-tagged anti-human-α-syn antibody for detection. (A) Measurement of pS 129 -α-syn in human CSF, serum, plasma, and saliva. HC, healthy control; PD, Parkinson’s disease; DLB, dementia with Lewy bodies; MSA, multiple system atrophy; LLoD, lower limit of detection; LLoQ, lower limit of quantitation. The p -values were calculated by a one-way ANOVA with post hoc Tukey test separately for the CSF samples and the serum, plasma, and saliva samples. (B) Comparison of pS 129 -α-syn concentrations in brain extracts from wild-type and MSA model mice. The p -values were calculated by Mann–Whitney test. The data are shown as the mean ± SD.

    Techniques Used: Quantitation Assay, MANN-WHITNEY

    Dilution linearity and spike recovery. (A–C) Dilution linearity within a 2-fold dilution series in (A) human CSF from a patient with DLB, (B) pooled human serum, and (C) extracts of PLP-α-syn mouse brain. The experiments were analyzed by repeated-measure one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. (D–F) Spike recovery rates after the addition of 250, 500, or 1000 pg/mL semisynthetic pS 129 -α-syn to (D) human CSF from a patient with DLB, (E) pooled human serum, and (F) extracts of PLP-α-syn mouse brain. The experiments were analyzed by ordinary one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. The data are shown as the mean ± SD.
    Figure Legend Snippet: Dilution linearity and spike recovery. (A–C) Dilution linearity within a 2-fold dilution series in (A) human CSF from a patient with DLB, (B) pooled human serum, and (C) extracts of PLP-α-syn mouse brain. The experiments were analyzed by repeated-measure one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. (D–F) Spike recovery rates after the addition of 250, 500, or 1000 pg/mL semisynthetic pS 129 -α-syn to (D) human CSF from a patient with DLB, (E) pooled human serum, and (F) extracts of PLP-α-syn mouse brain. The experiments were analyzed by ordinary one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. The data are shown as the mean ± SD.

    Techniques Used:

    Mass spectrometry analysis of the pS 129 -α-syn standard: an ESI mass-spectrum of pS 129 -α-syn. The inset shows the deconvoluted spectrum corresponding to a single protein species with the correct mass of pS 129 -α-syn.
    Figure Legend Snippet: Mass spectrometry analysis of the pS 129 -α-syn standard: an ESI mass-spectrum of pS 129 -α-syn. The inset shows the deconvoluted spectrum corresponding to a single protein species with the correct mass of pS 129 -α-syn.

    Techniques Used: Mass Spectrometry

    The pS 129 -α-syn standard aggregates when prepared using nonoptimized conditions. (A–D) Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows the presence of oligomers, presumably tetramers, and larger aggregates of pS 129 -α-syn but not of unphosphorylated α-syn. (B) Higher-sensitivity visualization by silver staining. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel. (E) 100 ng/mL pS 129 -α-syn was incubated at 37 °C for the indicated times, at which turbidity was measured as absorbance (scattering) at 600 nM and the electrochemiluminescence signal was measured as described in the .
    Figure Legend Snippet: The pS 129 -α-syn standard aggregates when prepared using nonoptimized conditions. (A–D) Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows the presence of oligomers, presumably tetramers, and larger aggregates of pS 129 -α-syn but not of unphosphorylated α-syn. (B) Higher-sensitivity visualization by silver staining. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel. (E) 100 ng/mL pS 129 -α-syn was incubated at 37 °C for the indicated times, at which turbidity was measured as absorbance (scattering) at 600 nM and the electrochemiluminescence signal was measured as described in the .

    Techniques Used: Staining, Silver Staining, Western Blot, Migration, Molecular Weight, Incubation, Electrochemiluminescence

    SDS–PAGE and Western-blot analysis of the pS 129 -α-syn standard prepared using optimized conditions. Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows an absence of oligomers of pS 129 -α-syn. (B) Higher-sensitivity visualization of the same gel by silver staining shows minor bands of a putative dimer in the 0.5 and 1.0 μg pS 129 -α-syn lanes. Minor degradation products are also observed under the monomer band. Bands between 50 and 60 kDa likely are keratin contamination and are not related to the analyzed proteins. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1 showing minor putative dimer bands in the 0.5 and 1.0 μg pS 129 -α-syn lanes. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel.
    Figure Legend Snippet: SDS–PAGE and Western-blot analysis of the pS 129 -α-syn standard prepared using optimized conditions. Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows an absence of oligomers of pS 129 -α-syn. (B) Higher-sensitivity visualization of the same gel by silver staining shows minor bands of a putative dimer in the 0.5 and 1.0 μg pS 129 -α-syn lanes. Minor degradation products are also observed under the monomer band. Bands between 50 and 60 kDa likely are keratin contamination and are not related to the analyzed proteins. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1 showing minor putative dimer bands in the 0.5 and 1.0 μg pS 129 -α-syn lanes. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel.

    Techniques Used: SDS Page, Western Blot, Staining, Silver Staining, Migration, Molecular Weight

    Standard curves for pS 129 -α-syn (red) and unphosphorylated α-syn standards (blue) using different antibody configurations. (A) Biotinylated EP1536Y used for capture and Sulfo-tagged MSD’s anti-human-α-syn antibody for detection. (B) Biotinylated MSD’s anti-human-α-syn antibody used for capture and Sulfo-tagged EP1536Y for detection. (C) Biotinylated mAb MJFR1 was used for capture and Sulfo-tagged EP1536Y for detection. Representative standard curves (mean ± SD of two technical replicates) of at least five independent experiments are shown.
    Figure Legend Snippet: Standard curves for pS 129 -α-syn (red) and unphosphorylated α-syn standards (blue) using different antibody configurations. (A) Biotinylated EP1536Y used for capture and Sulfo-tagged MSD’s anti-human-α-syn antibody for detection. (B) Biotinylated MSD’s anti-human-α-syn antibody used for capture and Sulfo-tagged EP1536Y for detection. (C) Biotinylated mAb MJFR1 was used for capture and Sulfo-tagged EP1536Y for detection. Representative standard curves (mean ± SD of two technical replicates) of at least five independent experiments are shown.

    Techniques Used:

    Evaluation of Antibody Combinations for Development of the  pS 129  -α-Synuclein Electrochemiluminescence ELISA <xref ref-type= a " title="Evaluation of Antibody Combinations for Development of the pS 129 -α-Synuclein Electrochemiluminescence ELISA
    Figure Legend Snippet: Evaluation of Antibody Combinations for Development of the pS 129 -α-Synuclein Electrochemiluminescence ELISA a

    Techniques Used: Electrochemiluminescence, Enzyme-linked Immunosorbent Assay, Intra Assay


    Structured Review

    Abcam igg ps beads
    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b <t>)</t> <t>BSA-PS</t> bead, ( c ) <t>IgG-PS</t> bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
    Igg Ps Beads, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    igg ps beads - by Bioz Stars, 2023-11
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    Images

    1) Product Images from "On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis"

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    Journal: Micromachines

    doi: 10.3390/mi14010206

    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
    Figure Legend Snippet: Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.

    Techniques Used: Concentration Assay

    Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.
    Figure Legend Snippet: Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.

    Techniques Used: Concentration Assay, Standard Deviation

    Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.
    Figure Legend Snippet: Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.

    Techniques Used:

    Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.
    Figure Legend Snippet: Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.

    Techniques Used:


    Structured Review

    Abcam igg bsa ps beads
    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) <t>IgG/BSA-PS</t> bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
    Igg Bsa Ps Beads, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis"

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    Journal: Micromachines

    doi: 10.3390/mi14010206

    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
    Figure Legend Snippet: Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.

    Techniques Used: Concentration Assay

    Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.
    Figure Legend Snippet: Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.

    Techniques Used: Concentration Assay, Standard Deviation

    Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.
    Figure Legend Snippet: Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.

    Techniques Used:

    Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.
    Figure Legend Snippet: Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.

    Techniques Used:


    Structured Review

    Abcam anti ps 473 akt
    Anti Ps 473 Akt, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti ps antibody 4b6
    Anti Ps Antibody 4b6, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Structured Review

    Abcam ps pt
    High pY levels induced by activation of cytokine and growth factor pathways were found in IPF lung tissues and fibroblasts. A , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 122) and control (n = 91) in dataset GSE47460. B , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 40) and control (n = 8) in dataset GSE53845. C , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 20) and control (n = 19) in dataset GSE92592. D-E , GSEA analysis in IPF dataset GSE47460. F-G , GSEA analysis in IPF dataset GSE71351. H , Representative images of Masson's trichrome <t>staining,</t> <t>α-SMA</t> and pY immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for α-SMA and pY. I , Representative images of <t>pS/pT</t> immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for pS/pT. J , Quantification analysis of immunostaining with α-SMA, pY and pS/T (n = 3). *** P < 0.001. K , Representative fluorescence images of pS/pT of lung tissue sections from different IPF patients and healthy control. L-M , Co-location analysis of pS/pT and α-SMA (n = 3). N , Representative fluorescence images of pY and α-SMA of lung tissue sections from IPF patients and healthy control. The boxed regions were revealed at higher magnification and arrows indicated overlap of pY and α-SMA in lung tissues. O-P , Co-location analysis of pY and α-SMA (n = 3). *** P < 0.001. Q-R , Western blot of pS/T and pY in hnLFs and hfLFs. GAPDH was used as a loading control. S , Western blot of FN, COL1A1 and α-SMA in lung fibroblasts from hnLFs and hfLFs. GAPDH was used as a loading control. T , Quantification of FN, COL1A1 and α-SMA protein levels in hnLFs and hfLFs (n = 3). * P < 0.05, *** P < 0.001. U , Representative fluorescence images of α-SMA and pY in hnLFs and hfLFs. V-W , Co-expression analysis of pS/pT and α-SMA (n = 3). *** P < 0.001.
    Ps Pt, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis"

    Article Title: Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis

    Journal: Theranostics

    doi: 10.7150/thno.72269

    High pY levels induced by activation of cytokine and growth factor pathways were found in IPF lung tissues and fibroblasts. A , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 122) and control (n = 91) in dataset GSE47460. B , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 40) and control (n = 8) in dataset GSE53845. C , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 20) and control (n = 19) in dataset GSE92592. D-E , GSEA analysis in IPF dataset GSE47460. F-G , GSEA analysis in IPF dataset GSE71351. H , Representative images of Masson's trichrome staining, α-SMA and pY immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for α-SMA and pY. I , Representative images of pS/pT immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for pS/pT. J , Quantification analysis of immunostaining with α-SMA, pY and pS/T (n = 3). *** P < 0.001. K , Representative fluorescence images of pS/pT of lung tissue sections from different IPF patients and healthy control. L-M , Co-location analysis of pS/pT and α-SMA (n = 3). N , Representative fluorescence images of pY and α-SMA of lung tissue sections from IPF patients and healthy control. The boxed regions were revealed at higher magnification and arrows indicated overlap of pY and α-SMA in lung tissues. O-P , Co-location analysis of pY and α-SMA (n = 3). *** P < 0.001. Q-R , Western blot of pS/T and pY in hnLFs and hfLFs. GAPDH was used as a loading control. S , Western blot of FN, COL1A1 and α-SMA in lung fibroblasts from hnLFs and hfLFs. GAPDH was used as a loading control. T , Quantification of FN, COL1A1 and α-SMA protein levels in hnLFs and hfLFs (n = 3). * P < 0.05, *** P < 0.001. U , Representative fluorescence images of α-SMA and pY in hnLFs and hfLFs. V-W , Co-expression analysis of pS/pT and α-SMA (n = 3). *** P < 0.001.
    Figure Legend Snippet: High pY levels induced by activation of cytokine and growth factor pathways were found in IPF lung tissues and fibroblasts. A , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 122) and control (n = 91) in dataset GSE47460. B , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 40) and control (n = 8) in dataset GSE53845. C , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 20) and control (n = 19) in dataset GSE92592. D-E , GSEA analysis in IPF dataset GSE47460. F-G , GSEA analysis in IPF dataset GSE71351. H , Representative images of Masson's trichrome staining, α-SMA and pY immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for α-SMA and pY. I , Representative images of pS/pT immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for pS/pT. J , Quantification analysis of immunostaining with α-SMA, pY and pS/T (n = 3). *** P < 0.001. K , Representative fluorescence images of pS/pT of lung tissue sections from different IPF patients and healthy control. L-M , Co-location analysis of pS/pT and α-SMA (n = 3). N , Representative fluorescence images of pY and α-SMA of lung tissue sections from IPF patients and healthy control. The boxed regions were revealed at higher magnification and arrows indicated overlap of pY and α-SMA in lung tissues. O-P , Co-location analysis of pY and α-SMA (n = 3). *** P < 0.001. Q-R , Western blot of pS/T and pY in hnLFs and hfLFs. GAPDH was used as a loading control. S , Western blot of FN, COL1A1 and α-SMA in lung fibroblasts from hnLFs and hfLFs. GAPDH was used as a loading control. T , Quantification of FN, COL1A1 and α-SMA protein levels in hnLFs and hfLFs (n = 3). * P < 0.05, *** P < 0.001. U , Representative fluorescence images of α-SMA and pY in hnLFs and hfLFs. V-W , Co-expression analysis of pS/pT and α-SMA (n = 3). *** P < 0.001.

    Techniques Used: Activation Assay, Staining, Immunostaining, Fluorescence, Western Blot, Expressing

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    Danaher Inc ps 1
    Experimental procedure and targeted molecular signaling. A Experimental protocol; B C3G-mediated amyloidogenic pathway and tau protein phosphorylation; C C3G enhanced autophagy in APPswe mice; D neuroprotective effects of C3G on neuronal apoptosis; and E C3G improved the synaptic function. APP, amyloid precursor protein; BACE1, β-secretase; <t>PS-1,</t> presenilin-1; ADAM10, α-secretase; Aβ, amyloid beta; IL, interleukin; TNF-α, tumor necrosis factor-α; GSH-Px, glutathione peroxidase; CAT, catalase; NFTs, neurofibrillary tangles; C3G, cyaninidin-3-O-glucoside; Bax, Bcl-2 associated X, apoptosis regulator; Bcl2, B-cell lymphoma 2; MAPKs, mitogen-activated protein kinases; GSK3β, glycogen synthase kinase-3 beta; AMPK, AMP-activated protein kinase; SIRT1, sirtuin 1; PI3K/Akt, phosphoinositide 3-kinase/protein kinase B; SYP, synaptophysin; PSD95, postsynaptic density protein-95; LC3B-II, light chain 3BII; LAMP-1, lysosomal-associated membrane protein 1; TFEB, transcription factor EB; SQSTM, sequestosome 1.
    Ps 1, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Danaher Inc antibody against pser129 ps α syn
    Experimental procedure and targeted molecular signaling. A Experimental protocol; B C3G-mediated amyloidogenic pathway and tau protein phosphorylation; C C3G enhanced autophagy in APPswe mice; D neuroprotective effects of C3G on neuronal apoptosis; and E C3G improved the synaptic function. APP, amyloid precursor protein; BACE1, β-secretase; <t>PS-1,</t> presenilin-1; ADAM10, α-secretase; Aβ, amyloid beta; IL, interleukin; TNF-α, tumor necrosis factor-α; GSH-Px, glutathione peroxidase; CAT, catalase; NFTs, neurofibrillary tangles; C3G, cyaninidin-3-O-glucoside; Bax, Bcl-2 associated X, apoptosis regulator; Bcl2, B-cell lymphoma 2; MAPKs, mitogen-activated protein kinases; GSK3β, glycogen synthase kinase-3 beta; AMPK, AMP-activated protein kinase; SIRT1, sirtuin 1; PI3K/Akt, phosphoinositide 3-kinase/protein kinase B; SYP, synaptophysin; PSD95, postsynaptic density protein-95; LC3B-II, light chain 3BII; LAMP-1, lysosomal-associated membrane protein 1; TFEB, transcription factor EB; SQSTM, sequestosome 1.
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    Abcam ps pkc substrate
    ( A ) Effects of various kinase inhibitors on the β CBP promoter via dual luciferase reporter assays. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells overexpressing ATF2 and incubated with the <t>PKC</t> inhibitor (20 μM), ERK inhibitor (30 μM), p38 inhibitor (30 μM), or JNK inhibitor (30 μM). DMSO was used as a control. P < 0.05, n = 3. ( B ) Relative levels of β CBP and ATF2 in the Pron of the fifth-instar gregarious locust nymphs injected with various inhibitors. Inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. * P < 0.05, n = 5 or 6. ( C ) Verification of ATF2 phosphorylation at serine stimulated by PKC. PKC inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. PBS was used as a control. ATF2 protein was enriched from the Pron of the fifth-instar gregarious locust nymphs using anti-ATF2 antibody-bound CNBr-activated Sepharose 4B. The phosphorylation of ATF2 was detected using anti–phospho-(Ser) PKC substrate (pS-PKC substrate) <t>polyclonal</t> <t>antibodies.</t> * P < 0.05 and ** P < 0.01, n = 4. ( D ) Expression is driven by the promoter of β CBP in the presence of various constructs. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells with ATF2 (WT), ATF2 S45A, ATF2 S327A, ATF2 S406A, ATF2 S518A, and ATF2 S668A, respectively. * P < 0.05, n = 3. Fluc/Rluc represents the fold of firefly to Renilla luciferase activity. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).
    Ps Pkc Substrate, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam rabbit anti ps
    ( A ) Effects of various kinase inhibitors on the β CBP promoter via dual luciferase reporter assays. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells overexpressing ATF2 and incubated with the <t>PKC</t> inhibitor (20 μM), ERK inhibitor (30 μM), p38 inhibitor (30 μM), or JNK inhibitor (30 μM). DMSO was used as a control. P < 0.05, n = 3. ( B ) Relative levels of β CBP and ATF2 in the Pron of the fifth-instar gregarious locust nymphs injected with various inhibitors. Inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. * P < 0.05, n = 5 or 6. ( C ) Verification of ATF2 phosphorylation at serine stimulated by PKC. PKC inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. PBS was used as a control. ATF2 protein was enriched from the Pron of the fifth-instar gregarious locust nymphs using anti-ATF2 antibody-bound CNBr-activated Sepharose 4B. The phosphorylation of ATF2 was detected using anti–phospho-(Ser) PKC substrate (pS-PKC substrate) <t>polyclonal</t> <t>antibodies.</t> * P < 0.05 and ** P < 0.01, n = 4. ( D ) Expression is driven by the promoter of β CBP in the presence of various constructs. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells with ATF2 (WT), ATF2 S45A, ATF2 S327A, ATF2 S406A, ATF2 S518A, and ATF2 S668A, respectively. * P < 0.05, n = 3. Fluc/Rluc represents the fold of firefly to Renilla luciferase activity. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).
    Rabbit Anti Ps, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti ps 129 α syn monoclonal antibody ep1536y
    Measurement of <t>pS</t> <t>129</t> <t>-α-syn</t> in human and mouse samples. All measurements were performed using biotinylated <t>EP1536Y</t> for capture and MSD’s Sulfo-tagged anti-human-α-syn antibody for detection. (A) Measurement of pS 129 -α-syn in human CSF, serum, plasma, and saliva. HC, healthy control; PD, Parkinson’s disease; DLB, dementia with Lewy bodies; MSA, multiple system atrophy; LLoD, lower limit of detection; LLoQ, lower limit of quantitation. The p -values were calculated by a one-way ANOVA with post hoc Tukey test separately for the CSF samples and the serum, plasma, and saliva samples. (B) Comparison of pS 129 -α-syn concentrations in brain extracts from wild-type and MSA model mice. The p -values were calculated by Mann–Whitney test. The data are shown as the mean ± SD.
    Anti Ps 129 α Syn Monoclonal Antibody Ep1536y, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam igg ps beads
    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b <t>)</t> <t>BSA-PS</t> bead, ( c ) <t>IgG-PS</t> bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
    Igg Ps Beads, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam igg bsa ps beads
    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) <t>IgG/BSA-PS</t> bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
    Igg Bsa Ps Beads, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti ps 473 akt
    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) <t>IgG/BSA-PS</t> bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
    Anti Ps 473 Akt, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam anti ps antibody 4b6
    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) <t>IgG/BSA-PS</t> bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.
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    ps pt  (Abcam)
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    Abcam ps pt
    High pY levels induced by activation of cytokine and growth factor pathways were found in IPF lung tissues and fibroblasts. A , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 122) and control (n = 91) in dataset GSE47460. B , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 40) and control (n = 8) in dataset GSE53845. C , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 20) and control (n = 19) in dataset GSE92592. D-E , GSEA analysis in IPF dataset GSE47460. F-G , GSEA analysis in IPF dataset GSE71351. H , Representative images of Masson's trichrome <t>staining,</t> <t>α-SMA</t> and pY immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for α-SMA and pY. I , Representative images of <t>pS/pT</t> immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for pS/pT. J , Quantification analysis of immunostaining with α-SMA, pY and pS/T (n = 3). *** P < 0.001. K , Representative fluorescence images of pS/pT of lung tissue sections from different IPF patients and healthy control. L-M , Co-location analysis of pS/pT and α-SMA (n = 3). N , Representative fluorescence images of pY and α-SMA of lung tissue sections from IPF patients and healthy control. The boxed regions were revealed at higher magnification and arrows indicated overlap of pY and α-SMA in lung tissues. O-P , Co-location analysis of pY and α-SMA (n = 3). *** P < 0.001. Q-R , Western blot of pS/T and pY in hnLFs and hfLFs. GAPDH was used as a loading control. S , Western blot of FN, COL1A1 and α-SMA in lung fibroblasts from hnLFs and hfLFs. GAPDH was used as a loading control. T , Quantification of FN, COL1A1 and α-SMA protein levels in hnLFs and hfLFs (n = 3). * P < 0.05, *** P < 0.001. U , Representative fluorescence images of α-SMA and pY in hnLFs and hfLFs. V-W , Co-expression analysis of pS/pT and α-SMA (n = 3). *** P < 0.001.
    Ps Pt, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Experimental procedure and targeted molecular signaling. A Experimental protocol; B C3G-mediated amyloidogenic pathway and tau protein phosphorylation; C C3G enhanced autophagy in APPswe mice; D neuroprotective effects of C3G on neuronal apoptosis; and E C3G improved the synaptic function. APP, amyloid precursor protein; BACE1, β-secretase; PS-1, presenilin-1; ADAM10, α-secretase; Aβ, amyloid beta; IL, interleukin; TNF-α, tumor necrosis factor-α; GSH-Px, glutathione peroxidase; CAT, catalase; NFTs, neurofibrillary tangles; C3G, cyaninidin-3-O-glucoside; Bax, Bcl-2 associated X, apoptosis regulator; Bcl2, B-cell lymphoma 2; MAPKs, mitogen-activated protein kinases; GSK3β, glycogen synthase kinase-3 beta; AMPK, AMP-activated protein kinase; SIRT1, sirtuin 1; PI3K/Akt, phosphoinositide 3-kinase/protein kinase B; SYP, synaptophysin; PSD95, postsynaptic density protein-95; LC3B-II, light chain 3BII; LAMP-1, lysosomal-associated membrane protein 1; TFEB, transcription factor EB; SQSTM, sequestosome 1.

    Journal: Journal of Neuroinflammation

    Article Title: Cyanidin-3- O -glucoside protects the brain and improves cognitive function in APPswe/PS1ΔE9 transgenic mice model

    doi: 10.1186/s12974-023-02950-3

    Figure Lengend Snippet: Experimental procedure and targeted molecular signaling. A Experimental protocol; B C3G-mediated amyloidogenic pathway and tau protein phosphorylation; C C3G enhanced autophagy in APPswe mice; D neuroprotective effects of C3G on neuronal apoptosis; and E C3G improved the synaptic function. APP, amyloid precursor protein; BACE1, β-secretase; PS-1, presenilin-1; ADAM10, α-secretase; Aβ, amyloid beta; IL, interleukin; TNF-α, tumor necrosis factor-α; GSH-Px, glutathione peroxidase; CAT, catalase; NFTs, neurofibrillary tangles; C3G, cyaninidin-3-O-glucoside; Bax, Bcl-2 associated X, apoptosis regulator; Bcl2, B-cell lymphoma 2; MAPKs, mitogen-activated protein kinases; GSK3β, glycogen synthase kinase-3 beta; AMPK, AMP-activated protein kinase; SIRT1, sirtuin 1; PI3K/Akt, phosphoinositide 3-kinase/protein kinase B; SYP, synaptophysin; PSD95, postsynaptic density protein-95; LC3B-II, light chain 3BII; LAMP-1, lysosomal-associated membrane protein 1; TFEB, transcription factor EB; SQSTM, sequestosome 1.

    Article Snippet: Antibodies against LC3B (ab48394), SQSTM1 (ab91526), TFEB (ab245350), LAMP-1 (ab24170), Bcl-2 (ab182858), Bax (ab32503), PPARα (ab24509), AMPK (ab3759), p-AMPK (T183) (ab133448), β-actin (ab6276), SIRT1 (ab110304), PS-1 (ab15458), APP (ab32136), BACE1 (ab183612), and ADAM10 (ab124695) were purchased from Abcam (Cambridge, UK).

    Techniques: Membrane

    ( A ) Effects of various kinase inhibitors on the β CBP promoter via dual luciferase reporter assays. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells overexpressing ATF2 and incubated with the PKC inhibitor (20 μM), ERK inhibitor (30 μM), p38 inhibitor (30 μM), or JNK inhibitor (30 μM). DMSO was used as a control. P < 0.05, n = 3. ( B ) Relative levels of β CBP and ATF2 in the Pron of the fifth-instar gregarious locust nymphs injected with various inhibitors. Inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. * P < 0.05, n = 5 or 6. ( C ) Verification of ATF2 phosphorylation at serine stimulated by PKC. PKC inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. PBS was used as a control. ATF2 protein was enriched from the Pron of the fifth-instar gregarious locust nymphs using anti-ATF2 antibody-bound CNBr-activated Sepharose 4B. The phosphorylation of ATF2 was detected using anti–phospho-(Ser) PKC substrate (pS-PKC substrate) polyclonal antibodies. * P < 0.05 and ** P < 0.01, n = 4. ( D ) Expression is driven by the promoter of β CBP in the presence of various constructs. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells with ATF2 (WT), ATF2 S45A, ATF2 S327A, ATF2 S406A, ATF2 S518A, and ATF2 S668A, respectively. * P < 0.05, n = 3. Fluc/Rluc represents the fold of firefly to Renilla luciferase activity. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).

    Journal: Science Advances

    Article Title: Spatially differential regulation of ATF2 phosphorylation contributes to warning coloration of gregarious locusts

    doi: 10.1126/sciadv.adi5168

    Figure Lengend Snippet: ( A ) Effects of various kinase inhibitors on the β CBP promoter via dual luciferase reporter assays. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells overexpressing ATF2 and incubated with the PKC inhibitor (20 μM), ERK inhibitor (30 μM), p38 inhibitor (30 μM), or JNK inhibitor (30 μM). DMSO was used as a control. P < 0.05, n = 3. ( B ) Relative levels of β CBP and ATF2 in the Pron of the fifth-instar gregarious locust nymphs injected with various inhibitors. Inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. * P < 0.05, n = 5 or 6. ( C ) Verification of ATF2 phosphorylation at serine stimulated by PKC. PKC inhibitor (50 μM per nymph) was injected into the mid-third instar gregarious locust nymphs twice over 48-hour intervals. PBS was used as a control. ATF2 protein was enriched from the Pron of the fifth-instar gregarious locust nymphs using anti-ATF2 antibody-bound CNBr-activated Sepharose 4B. The phosphorylation of ATF2 was detected using anti–phospho-(Ser) PKC substrate (pS-PKC substrate) polyclonal antibodies. * P < 0.05 and ** P < 0.01, n = 4. ( D ) Expression is driven by the promoter of β CBP in the presence of various constructs. pGL4.10-β CBP −184 to +1 and pGL4.73 were cotransfected into S2 cells with ATF2 (WT), ATF2 S45A, ATF2 S327A, ATF2 S406A, ATF2 S518A, and ATF2 S668A, respectively. * P < 0.05, n = 3. Fluc/Rluc represents the fold of firefly to Renilla luciferase activity. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).

    Article Snippet: Antibodies against p-Ser/Thr, pS-PKC substrate, and p-Thr were purchased from Abcam and Cell Signaling Technology.

    Techniques: Luciferase, Incubation, Injection, Expressing, Construct, Activity Assay, Two Tailed Test

    ( A ) β CBP and ATF2 mRNA levels after PKC α knockdown in Pron. PKC α knockdown (3 μg per nymph administered in the day 2 of the second-instar gregarious locust nymphs, four times over 48-hours intervals). dsGFP was used as a control. ** P < 0.01, n = 4. ( B ) ATF2 phosphorylation levels in Pron after PKC isoform knockdown. ** P < 0.01, n = 3 or 4. ( C ) Body color phenotypes of gregarious locust nymphs after RNAi exposure. Images were obtained on the fifth-instar nymphs after RNAi exposure. Red box represents locusts with a notable black-brown pattern. Blue box represents locusts with a cryptic green color. Orange box represents locusts with a body color that falls between the notable black-brown pattern and the cryptic green color. Scale bar, 0.5 cm. * P < 0.05 and ** P < 0.01, n = 3. ( D ) Immunohistochemistry of ATF2 and pS-ATF2 in Pros after PKC α knockdown. Green denotes ATF2 or pS-ATF2 staining by anti-ATF2 antibodies or anti–phospho-ATF2 (Ser 327 ) antibodies. Blue denotes DAPI staining for nuclei. Scale bars, 25 μm. ( E ) β-carotene content coupled with βCBP was evaluated in Pron after PKC α knockdown by HPLC. * P < 0.05, n = 5. ( F ) ChIP assays show that PKC α knockdown repressed the relative precipitation of the β CBP promoter region in Pros. pS-ATF2, phospho-ATF2 (Ser 327 ) antibodies; ATF2, ATF2 antibodies; IgG, nonspecific rabbit IgG control. P < 0.05, n = 3 or 4. Pron: The fifth-instar gregarious locust nymphs pronotum. Pros: The fifth-instar gregarious locust nymphs prosternum. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).

    Journal: Science Advances

    Article Title: Spatially differential regulation of ATF2 phosphorylation contributes to warning coloration of gregarious locusts

    doi: 10.1126/sciadv.adi5168

    Figure Lengend Snippet: ( A ) β CBP and ATF2 mRNA levels after PKC α knockdown in Pron. PKC α knockdown (3 μg per nymph administered in the day 2 of the second-instar gregarious locust nymphs, four times over 48-hours intervals). dsGFP was used as a control. ** P < 0.01, n = 4. ( B ) ATF2 phosphorylation levels in Pron after PKC isoform knockdown. ** P < 0.01, n = 3 or 4. ( C ) Body color phenotypes of gregarious locust nymphs after RNAi exposure. Images were obtained on the fifth-instar nymphs after RNAi exposure. Red box represents locusts with a notable black-brown pattern. Blue box represents locusts with a cryptic green color. Orange box represents locusts with a body color that falls between the notable black-brown pattern and the cryptic green color. Scale bar, 0.5 cm. * P < 0.05 and ** P < 0.01, n = 3. ( D ) Immunohistochemistry of ATF2 and pS-ATF2 in Pros after PKC α knockdown. Green denotes ATF2 or pS-ATF2 staining by anti-ATF2 antibodies or anti–phospho-ATF2 (Ser 327 ) antibodies. Blue denotes DAPI staining for nuclei. Scale bars, 25 μm. ( E ) β-carotene content coupled with βCBP was evaluated in Pron after PKC α knockdown by HPLC. * P < 0.05, n = 5. ( F ) ChIP assays show that PKC α knockdown repressed the relative precipitation of the β CBP promoter region in Pros. pS-ATF2, phospho-ATF2 (Ser 327 ) antibodies; ATF2, ATF2 antibodies; IgG, nonspecific rabbit IgG control. P < 0.05, n = 3 or 4. Pron: The fifth-instar gregarious locust nymphs pronotum. Pros: The fifth-instar gregarious locust nymphs prosternum. Data are presented as the means ± SDs. Asterisks indicate statistically significant differences based on two-tailed Student’s t tests (* P < 0.05 and ** P < 0.01). Different letters indicate multiple comparisons of significant differences based on one-way ANOVA followed by Tukey’s test ( P < 0.05).

    Article Snippet: Antibodies against p-Ser/Thr, pS-PKC substrate, and p-Thr were purchased from Abcam and Cell Signaling Technology.

    Techniques: Immunohistochemistry, Staining, Two Tailed Test

    Measurement of pS 129 -α-syn in human and mouse samples. All measurements were performed using biotinylated EP1536Y for capture and MSD’s Sulfo-tagged anti-human-α-syn antibody for detection. (A) Measurement of pS 129 -α-syn in human CSF, serum, plasma, and saliva. HC, healthy control; PD, Parkinson’s disease; DLB, dementia with Lewy bodies; MSA, multiple system atrophy; LLoD, lower limit of detection; LLoQ, lower limit of quantitation. The p -values were calculated by a one-way ANOVA with post hoc Tukey test separately for the CSF samples and the serum, plasma, and saliva samples. (B) Comparison of pS 129 -α-syn concentrations in brain extracts from wild-type and MSA model mice. The p -values were calculated by Mann–Whitney test. The data are shown as the mean ± SD.

    Journal: ACS Chemical Neuroscience

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    doi: 10.1021/acschemneuro.2c00676

    Figure Lengend Snippet: Measurement of pS 129 -α-syn in human and mouse samples. All measurements were performed using biotinylated EP1536Y for capture and MSD’s Sulfo-tagged anti-human-α-syn antibody for detection. (A) Measurement of pS 129 -α-syn in human CSF, serum, plasma, and saliva. HC, healthy control; PD, Parkinson’s disease; DLB, dementia with Lewy bodies; MSA, multiple system atrophy; LLoD, lower limit of detection; LLoQ, lower limit of quantitation. The p -values were calculated by a one-way ANOVA with post hoc Tukey test separately for the CSF samples and the serum, plasma, and saliva samples. (B) Comparison of pS 129 -α-syn concentrations in brain extracts from wild-type and MSA model mice. The p -values were calculated by Mann–Whitney test. The data are shown as the mean ± SD.

    Article Snippet: Anti-pS 129 -α-syn monoclonal antibody EP1536Y (ab209422) and anti-α-synuclein monoclonal antibody MJFR1 (ab138501) were procured from Abcam (Cambridge, UK).

    Techniques: Quantitation Assay, MANN-WHITNEY

    Dilution linearity and spike recovery. (A–C) Dilution linearity within a 2-fold dilution series in (A) human CSF from a patient with DLB, (B) pooled human serum, and (C) extracts of PLP-α-syn mouse brain. The experiments were analyzed by repeated-measure one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. (D–F) Spike recovery rates after the addition of 250, 500, or 1000 pg/mL semisynthetic pS 129 -α-syn to (D) human CSF from a patient with DLB, (E) pooled human serum, and (F) extracts of PLP-α-syn mouse brain. The experiments were analyzed by ordinary one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. The data are shown as the mean ± SD.

    Journal: ACS Chemical Neuroscience

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    doi: 10.1021/acschemneuro.2c00676

    Figure Lengend Snippet: Dilution linearity and spike recovery. (A–C) Dilution linearity within a 2-fold dilution series in (A) human CSF from a patient with DLB, (B) pooled human serum, and (C) extracts of PLP-α-syn mouse brain. The experiments were analyzed by repeated-measure one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. (D–F) Spike recovery rates after the addition of 250, 500, or 1000 pg/mL semisynthetic pS 129 -α-syn to (D) human CSF from a patient with DLB, (E) pooled human serum, and (F) extracts of PLP-α-syn mouse brain. The experiments were analyzed by ordinary one-way ANOVA with Tukey’s multiple comparisons test with a single pooled variance. The data are shown as the mean ± SD.

    Article Snippet: Anti-pS 129 -α-syn monoclonal antibody EP1536Y (ab209422) and anti-α-synuclein monoclonal antibody MJFR1 (ab138501) were procured from Abcam (Cambridge, UK).

    Techniques:

    Mass spectrometry analysis of the pS 129 -α-syn standard: an ESI mass-spectrum of pS 129 -α-syn. The inset shows the deconvoluted spectrum corresponding to a single protein species with the correct mass of pS 129 -α-syn.

    Journal: ACS Chemical Neuroscience

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    doi: 10.1021/acschemneuro.2c00676

    Figure Lengend Snippet: Mass spectrometry analysis of the pS 129 -α-syn standard: an ESI mass-spectrum of pS 129 -α-syn. The inset shows the deconvoluted spectrum corresponding to a single protein species with the correct mass of pS 129 -α-syn.

    Article Snippet: Anti-pS 129 -α-syn monoclonal antibody EP1536Y (ab209422) and anti-α-synuclein monoclonal antibody MJFR1 (ab138501) were procured from Abcam (Cambridge, UK).

    Techniques: Mass Spectrometry

    The pS 129 -α-syn standard aggregates when prepared using nonoptimized conditions. (A–D) Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows the presence of oligomers, presumably tetramers, and larger aggregates of pS 129 -α-syn but not of unphosphorylated α-syn. (B) Higher-sensitivity visualization by silver staining. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel. (E) 100 ng/mL pS 129 -α-syn was incubated at 37 °C for the indicated times, at which turbidity was measured as absorbance (scattering) at 600 nM and the electrochemiluminescence signal was measured as described in the .

    Journal: ACS Chemical Neuroscience

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    doi: 10.1021/acschemneuro.2c00676

    Figure Lengend Snippet: The pS 129 -α-syn standard aggregates when prepared using nonoptimized conditions. (A–D) Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows the presence of oligomers, presumably tetramers, and larger aggregates of pS 129 -α-syn but not of unphosphorylated α-syn. (B) Higher-sensitivity visualization by silver staining. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel. (E) 100 ng/mL pS 129 -α-syn was incubated at 37 °C for the indicated times, at which turbidity was measured as absorbance (scattering) at 600 nM and the electrochemiluminescence signal was measured as described in the .

    Article Snippet: Anti-pS 129 -α-syn monoclonal antibody EP1536Y (ab209422) and anti-α-synuclein monoclonal antibody MJFR1 (ab138501) were procured from Abcam (Cambridge, UK).

    Techniques: Staining, Silver Staining, Western Blot, Migration, Molecular Weight, Incubation, Electrochemiluminescence

    SDS–PAGE and Western-blot analysis of the pS 129 -α-syn standard prepared using optimized conditions. Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows an absence of oligomers of pS 129 -α-syn. (B) Higher-sensitivity visualization of the same gel by silver staining shows minor bands of a putative dimer in the 0.5 and 1.0 μg pS 129 -α-syn lanes. Minor degradation products are also observed under the monomer band. Bands between 50 and 60 kDa likely are keratin contamination and are not related to the analyzed proteins. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1 showing minor putative dimer bands in the 0.5 and 1.0 μg pS 129 -α-syn lanes. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel.

    Journal: ACS Chemical Neuroscience

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    doi: 10.1021/acschemneuro.2c00676

    Figure Lengend Snippet: SDS–PAGE and Western-blot analysis of the pS 129 -α-syn standard prepared using optimized conditions. Different quantities of the semisynthetic pS 129 -α-syn standard were fractionated, and unphosphorylated α-syn was used as a control. (A) Coomassie Blue staining shows an absence of oligomers of pS 129 -α-syn. (B) Higher-sensitivity visualization of the same gel by silver staining shows minor bands of a putative dimer in the 0.5 and 1.0 μg pS 129 -α-syn lanes. Minor degradation products are also observed under the monomer band. Bands between 50 and 60 kDa likely are keratin contamination and are not related to the analyzed proteins. (C) Western blot analysis probed with the anti-α-syn antibody MJFR1 showing minor putative dimer bands in the 0.5 and 1.0 μg pS 129 -α-syn lanes. (D) Western blot analysis probed with the anti-pS 129 -α-syn antibody EP1536Y. The gel migration of molecular weight markers is shown on the left in each panel.

    Article Snippet: Anti-pS 129 -α-syn monoclonal antibody EP1536Y (ab209422) and anti-α-synuclein monoclonal antibody MJFR1 (ab138501) were procured from Abcam (Cambridge, UK).

    Techniques: SDS Page, Western Blot, Staining, Silver Staining, Migration, Molecular Weight

    Standard curves for pS 129 -α-syn (red) and unphosphorylated α-syn standards (blue) using different antibody configurations. (A) Biotinylated EP1536Y used for capture and Sulfo-tagged MSD’s anti-human-α-syn antibody for detection. (B) Biotinylated MSD’s anti-human-α-syn antibody used for capture and Sulfo-tagged EP1536Y for detection. (C) Biotinylated mAb MJFR1 was used for capture and Sulfo-tagged EP1536Y for detection. Representative standard curves (mean ± SD of two technical replicates) of at least five independent experiments are shown.

    Journal: ACS Chemical Neuroscience

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    doi: 10.1021/acschemneuro.2c00676

    Figure Lengend Snippet: Standard curves for pS 129 -α-syn (red) and unphosphorylated α-syn standards (blue) using different antibody configurations. (A) Biotinylated EP1536Y used for capture and Sulfo-tagged MSD’s anti-human-α-syn antibody for detection. (B) Biotinylated MSD’s anti-human-α-syn antibody used for capture and Sulfo-tagged EP1536Y for detection. (C) Biotinylated mAb MJFR1 was used for capture and Sulfo-tagged EP1536Y for detection. Representative standard curves (mean ± SD of two technical replicates) of at least five independent experiments are shown.

    Article Snippet: Anti-pS 129 -α-syn monoclonal antibody EP1536Y (ab209422) and anti-α-synuclein monoclonal antibody MJFR1 (ab138501) were procured from Abcam (Cambridge, UK).

    Techniques:

    Evaluation of Antibody Combinations for Development of the  pS 129  -α-Synuclein Electrochemiluminescence ELISA <xref ref-type= a " width="100%" height="100%">

    Journal: ACS Chemical Neuroscience

    Article Title: Development of a Novel Electrochemiluminescence ELISA for Quantification of α-Synuclein Phosphorylated at Ser 129 in Biological Samples

    doi: 10.1021/acschemneuro.2c00676

    Figure Lengend Snippet: Evaluation of Antibody Combinations for Development of the pS 129 -α-Synuclein Electrochemiluminescence ELISA a

    Article Snippet: Anti-pS 129 -α-syn monoclonal antibody EP1536Y (ab209422) and anti-α-synuclein monoclonal antibody MJFR1 (ab138501) were procured from Abcam (Cambridge, UK).

    Techniques: Electrochemiluminescence, Enzyme-linked Immunosorbent Assay, Intra Assay

    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques: Concentration Assay

    Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques: Concentration Assay, Standard Deviation

    Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques:

    Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques:

    Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Four kinds of antigen bead and measurement methods for antigen bead and microneedle phagocytosis . ( A ). Schematic drawings of samples: ( a ) polystyrene (PS) bead, ( b ) BSA-PS bead, ( c ) IgG-PS bead, ( d ) IgG/BSA-PS bead. ( B ). Procedure of antigen bead phagocytosis experiment by the on-chip free-flow method. Antigen beads were flown in the dish with constant speed to keep the concentration of surrounding macrophages constant. ( C ). Experimental setup for free-flow method. Since macrophages adhered to the bottom of the dish, tilting the dish could generate a flow of antigen beads to the macrophages. ( D ). Experimental setup of bead feeding with optical tweezers. To feed antigen beads one-by-one to macrophages, the beads were trapped and transferred with optical tweezers. ( E ). Experimental setup for IgG/BSA-coated microneedle phagocytosis. The tip of the antigen microneedles was brought into contact with the macrophages by moving the motorized manipulator to which the antigen microneedle was attached.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques: Concentration Assay

    Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Time-course change in antigen concentration and total number of phagocytozed macrophages in the on-chip free-flow method. ( A ). Time-course change in antigen bead concentration and the total number of antigen beads phagocytozed by macrophages. ( a ) Time-course change in flowing antigen concentration at the observation area. The concentration of flowing antigens became stable 70 min after the flow started. The black dashed line is the model line when the antigen concentration is assumed to increase at a constant rate during the initial 70 min toward the bead concentration after 70 min (635 particles/mm 2 ) and then remain constant. ( b ) Total number increase of antigens phagocytozed by 33 macrophages. The black dashed line is the fitting line using the least-squares method for the plot after 40 min when the rate of increase in the number of phagocytozed beads became constant. The slope of the dashed line is 1.5/min. ( c ) Micrograph of the entire observation area at 70 min when the antigen concentration became constant. Bar: 100 μ m . ( d ) Micrograph of macrophages phagocytosing antigen beads. The m in the lower-left of each picture represents the engulfed antigen number. Bar: 10 μ m . ( B ). Sample surface decoration dependence of phagocytic efficiency. The phagocytic efficiency of five types of antigens was compared: IgG/BSA-PS (light blue dots, error bars and line, n = 100 cells), IgG-PS (gray, n = 100), BSA-PS (orange, n = 100), PS (dark blue, n = 100), and Zymosan (green, n = 40). The dots and error bars represent the mean and standard deviation of the experimental values, respectively, and the lines are the fitting curves according to Equation . The phagocytosis ratio at each time represents the percentage of macrophages that phagocytozed at least one antigen relative to the total number of macrophages. Since Zymosan is a digestible biological sample, data for 1440 min could not be obtained because it is not possible to confirm by direct observation whether or not the macrophages after an extended period are cells after phagocytosis.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques: Concentration Assay, Standard Deviation

    Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Evaluation of phagocytic antigen number dependence of phagocytosis time after feeding with optical tweezers. ( A ). Schematic drawing and micrographs of one-by-one phagocytosis process. ( a ) Transport of first antigen IgG/BSA-PS bead with optical tweezers. ( b ) Contact of first antigen bead with cell. ( c ) Removal of optical trapping after antigen bead adhesion. ( d ) Completion of phagocytosis for first bead. ( e ) Contact of second antigen bead. ( f ) Contact of antigen bead after the macrophage reached its maximum limit of phagocytosis. ( g – i ) Contact of additional beads on the other areas of the cell. ( j ) Final state. Bar: 10 μ m . ( B ). Typical example of phagocytosis number dependence of phagocytosis time in 2 μ m ( a ) and 4.5 μ m ( b ) IgG/BSA-PS bead phagocytosis, respectively. ( C ). Histogram of difference in phagocytosis time in adjacent phagocytosis in 2 μ m (( a ), n = 235 phagocytosis) and 4.5 μ m (( b ), n = 79) IgG/BSA-PS bead phagocytosis, respectively.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques:

    Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.

    Journal: Micromachines

    Article Title: On-Chip Free-Flow Measurement Revealed Possible Depletion of Macrophages by Indigestible PM2.5 within a Few Hours by the Fastest Intervals of Serial Phagocytosis

    doi: 10.3390/mi14010206

    Figure Lengend Snippet: Waiting time from antigen contact to the start of phagocytosis and progress speed in IgG/BSA-coated microneedle phagocytosis. ( A ). Evaluation of waiting time from antigen microneedle contact to the start of phagocytosis in two-microneedle phagocytosis (n = 21 samples). ( a ) Histogram of waiting time. The time from microneedle contact to the start of phagocytosis was measured for each of the two needles. ( b ) Difference in waiting time in the first and second needles. ( B ). Typical example of cell membrane progression on the first (red) and second (blue) microneedles in two-microneedle phagocytosis. The edges of the cell membrane on the first and second microneedles are indicated by red and blue arrowheads in the micrographs. Bar: 10 μ m . ( C ). Evaluation of the difference in the progress speed of the cell membrane on the first and second microneedles. The progress speed was measured for each of the two microneedles, and the ratio of these differences to the progress speed on the first needle, i.e., 100 × ((2nd) − (1st))/(1st), was calculated.

    Article Snippet: For confirmation of the IgG decoration on the surface of IgG-PS beads, IgG/BSA-PS beads, and IgG/BSA-glass microneedles, a secondary antibody against IgG with Alexa Fluor 488 (Abcam, Cambridge, UK) was used and observed with a fluorescent invert optical microscope (IX71; Olympus, Tokyo, Japan).

    Techniques:

    High pY levels induced by activation of cytokine and growth factor pathways were found in IPF lung tissues and fibroblasts. A , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 122) and control (n = 91) in dataset GSE47460. B , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 40) and control (n = 8) in dataset GSE53845. C , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 20) and control (n = 19) in dataset GSE92592. D-E , GSEA analysis in IPF dataset GSE47460. F-G , GSEA analysis in IPF dataset GSE71351. H , Representative images of Masson's trichrome staining, α-SMA and pY immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for α-SMA and pY. I , Representative images of pS/pT immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for pS/pT. J , Quantification analysis of immunostaining with α-SMA, pY and pS/T (n = 3). *** P < 0.001. K , Representative fluorescence images of pS/pT of lung tissue sections from different IPF patients and healthy control. L-M , Co-location analysis of pS/pT and α-SMA (n = 3). N , Representative fluorescence images of pY and α-SMA of lung tissue sections from IPF patients and healthy control. The boxed regions were revealed at higher magnification and arrows indicated overlap of pY and α-SMA in lung tissues. O-P , Co-location analysis of pY and α-SMA (n = 3). *** P < 0.001. Q-R , Western blot of pS/T and pY in hnLFs and hfLFs. GAPDH was used as a loading control. S , Western blot of FN, COL1A1 and α-SMA in lung fibroblasts from hnLFs and hfLFs. GAPDH was used as a loading control. T , Quantification of FN, COL1A1 and α-SMA protein levels in hnLFs and hfLFs (n = 3). * P < 0.05, *** P < 0.001. U , Representative fluorescence images of α-SMA and pY in hnLFs and hfLFs. V-W , Co-expression analysis of pS/pT and α-SMA (n = 3). *** P < 0.001.

    Journal: Theranostics

    Article Title: Blockade of phosphotyrosine pathways suggesting SH2 superbinder as a novel therapy for pulmonary fibrosis

    doi: 10.7150/thno.72269

    Figure Lengend Snippet: High pY levels induced by activation of cytokine and growth factor pathways were found in IPF lung tissues and fibroblasts. A , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 122) and control (n = 91) in dataset GSE47460. B , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 40) and control (n = 8) in dataset GSE53845. C , KEGG pathway enrichment analysis in lung tissues from IPF patients (n = 20) and control (n = 19) in dataset GSE92592. D-E , GSEA analysis in IPF dataset GSE47460. F-G , GSEA analysis in IPF dataset GSE71351. H , Representative images of Masson's trichrome staining, α-SMA and pY immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for α-SMA and pY. I , Representative images of pS/pT immunostaining of lung tissue sections from IPF patients and healthy control. IgG was an isotype control for pS/pT. J , Quantification analysis of immunostaining with α-SMA, pY and pS/T (n = 3). *** P < 0.001. K , Representative fluorescence images of pS/pT of lung tissue sections from different IPF patients and healthy control. L-M , Co-location analysis of pS/pT and α-SMA (n = 3). N , Representative fluorescence images of pY and α-SMA of lung tissue sections from IPF patients and healthy control. The boxed regions were revealed at higher magnification and arrows indicated overlap of pY and α-SMA in lung tissues. O-P , Co-location analysis of pY and α-SMA (n = 3). *** P < 0.001. Q-R , Western blot of pS/T and pY in hnLFs and hfLFs. GAPDH was used as a loading control. S , Western blot of FN, COL1A1 and α-SMA in lung fibroblasts from hnLFs and hfLFs. GAPDH was used as a loading control. T , Quantification of FN, COL1A1 and α-SMA protein levels in hnLFs and hfLFs (n = 3). * P < 0.05, *** P < 0.001. U , Representative fluorescence images of α-SMA and pY in hnLFs and hfLFs. V-W , Co-expression analysis of pS/pT and α-SMA (n = 3). *** P < 0.001.

    Article Snippet: Antibodies against pY (Cat#EPR16871), p-PDGFRβ (Cat#ab248657), p-FGFR1 (Cat#ab59194), α-SMA (Cat#ab124964), pS/pT (Cat#ab117253) and IgG (Cat#EPR25A and Cat#ab37415) were purchased from Abcam (Cambridge, MA, USA).

    Techniques: Activation Assay, Staining, Immunostaining, Fluorescence, Western Blot, Expressing