Review



pip3 indicator pcdna3 1 aktph mcherry  (Addgene inc)


Bioz Verified Symbol Addgene inc is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 93
      Buy from Supplier

    Structured Review

    Addgene inc pip3 indicator pcdna3 1 aktph mcherry
    (a) Representative images of 3T3 cells co-transfected with <t>PIP3</t> indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.
    Pip3 Indicator Pcdna3 1 Aktph Mcherry, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pip3 indicator pcdna3 1 aktph mcherry/product/Addgene inc
    Average 93 stars, based on 8 article reviews
    pip3 indicator pcdna3 1 aktph mcherry - by Bioz Stars, 2026-02
    93/100 stars

    Images

    1) Product Images from "Chemical Compensation to Mechanical Loss in Cell Mechanosensation"

    Article Title: Chemical Compensation to Mechanical Loss in Cell Mechanosensation

    Journal: bioRxiv

    doi: 10.1101/2025.04.02.646640

    (a) Representative images of 3T3 cells co-transfected with PIP3 indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.
    Figure Legend Snippet: (a) Representative images of 3T3 cells co-transfected with PIP3 indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.

    Techniques Used: Transfection, Membrane, Control, Comparison, Western Blot, Activity Assay, MANN-WHITNEY

    (a-b) Normalized PIP3 dynamics of 3T3 cells treated with 10 µM LY29 (a) and 20 µM Y-27 (b). PIP3 level is quantified by the ratio of the PIP3 indicator (AktPH-mCherry) and the membrane indicator (GFP-CaaX) at the cell boundary. (c) Representative western blot images and quantifications of Akt activity in 3T3 cells, treated with vehicle, 2 µM LatA, and 20 µM Y-27 for 30 min. (d) Representative western blot images and quantifications of Akt activity in 3T3 and HT1080 cells treated with vehicle or 40 µM EIPA. (e) Chemical module activity of 3T3 versus HT1080 predicted by the model. (f,g) NHE1 (f) and the chemical module (g) activities modeled by reducing , or both. Only by reducing both parameters was able to reproduce the insensitive response in HT1080. (h) Volume dynamics of 3T3 Piezo1 KO cells treated with 0.2 µM Trametinib (Trame) with hypotonic shock applied at 30 min. (e-g) 3T3 parameters are set as: , n 12 = n 32 = n 13 = n 23 = ™ 2, R 12 = R 23 = R 31 = 1 / 5. HT1080 parameters are the same as 3T3 excepted for and . All results are normalized to a base activity level at . (a-c, h) Error bars represent SEM. (d) Error bars represent STD.
    Figure Legend Snippet: (a-b) Normalized PIP3 dynamics of 3T3 cells treated with 10 µM LY29 (a) and 20 µM Y-27 (b). PIP3 level is quantified by the ratio of the PIP3 indicator (AktPH-mCherry) and the membrane indicator (GFP-CaaX) at the cell boundary. (c) Representative western blot images and quantifications of Akt activity in 3T3 cells, treated with vehicle, 2 µM LatA, and 20 µM Y-27 for 30 min. (d) Representative western blot images and quantifications of Akt activity in 3T3 and HT1080 cells treated with vehicle or 40 µM EIPA. (e) Chemical module activity of 3T3 versus HT1080 predicted by the model. (f,g) NHE1 (f) and the chemical module (g) activities modeled by reducing , or both. Only by reducing both parameters was able to reproduce the insensitive response in HT1080. (h) Volume dynamics of 3T3 Piezo1 KO cells treated with 0.2 µM Trametinib (Trame) with hypotonic shock applied at 30 min. (e-g) 3T3 parameters are set as: , n 12 = n 32 = n 13 = n 23 = ™ 2, R 12 = R 23 = R 31 = 1 / 5. HT1080 parameters are the same as 3T3 excepted for and . All results are normalized to a base activity level at . (a-c, h) Error bars represent SEM. (d) Error bars represent STD.

    Techniques Used: Membrane, Western Blot, Activity Assay



    Similar Products

    pip3 indicator pcdna3 1 aktph mcherry  (Addgene inc)
    93
    Addgene inc pip3 indicator pcdna3 1 aktph mcherry
    (a) Representative images of 3T3 cells co-transfected with <t>PIP3</t> indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.
    Pip3 Indicator Pcdna3 1 Aktph Mcherry, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pip3 indicator pcdna3 1 aktph mcherry/product/Addgene inc
    Average 93 stars, based on 1 article reviews
    pip3 indicator pcdna3 1 aktph mcherry - by Bioz Stars, 2026-02
    93/100 stars
    		  Buy from Supplier
    aktph pmcherry 92  (Addgene inc)
    93
    Addgene inc aktph pmcherry 92
    (a) Representative images of 3T3 cells co-transfected with <t>PIP3</t> indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.
    Aktph Pmcherry 92, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aktph pmcherry 92/product/Addgene inc
    Average 93 stars, based on 1 article reviews
    aktph pmcherry 92 - by Bioz Stars, 2026-02
    93/100 stars
    		  Buy from Supplier
    aktph pmcherry  (Addgene inc)
    93
    Addgene inc aktph pmcherry
    (a) Representative images of 3T3 cells co-transfected with <t>PIP3</t> indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.
    Aktph Pmcherry, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/aktph pmcherry/product/Addgene inc
    Average 93 stars, based on 1 article reviews
    aktph pmcherry - by Bioz Stars, 2026-02
    93/100 stars
    		  Buy from Supplier
    pcdna3 1 aktph mcherry  (Addgene inc)
    93
    Addgene inc pcdna3 1 aktph mcherry
    (a) Representative images of 3T3 cells co-transfected with <t>PIP3</t> indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.
    Pcdna3 1 Aktph Mcherry, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcdna3 1 aktph mcherry/product/Addgene inc
    Average 93 stars, based on 1 article reviews
    pcdna3 1 aktph mcherry - by Bioz Stars, 2026-02
    93/100 stars
    		  Buy from Supplier

    Image Search Results


    (a) Representative images of 3T3 cells co-transfected with PIP3 indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.

    Journal: bioRxiv

    Article Title: Chemical Compensation to Mechanical Loss in Cell Mechanosensation

    doi: 10.1101/2025.04.02.646640

    Figure Lengend Snippet: (a) Representative images of 3T3 cells co-transfected with PIP3 indicator (AktPH-mCherry) and membrane indicator (GFP-CaaX), and their ratio at the cell boundary. The ratio at the cell boundary indicates the relative PIP3 level. (b) Normalized PIP3 dynamics of 3T3 cells treated with vehicle control and 2 µM LatA. (c) Normalized PIP3 level of 3T3 treated with vehicle and 2 µM LatA. n = 30, 21, 18, 31. (d) ΔpH i of 3T3-pHred cells treated with vehicle and 10 µM LY294002 (LY29) for 1 h. n = 36, 28. (e) ΔpH i of 3T3-pHred cells pre-treated with 10 µM LY29 for 2 h prior to treatment of vehicle, 2 µM LatA, and 20 µM Y-27. n = 39, 28, 47. (f) Comparison of the 3T3 volume dynamics treated with vehicle, 5 µM n-Bleb, and a combination of 5 µM n-Bleb and 10 µM LY29, with hypotonic shock applied at 30 min. (g) Representative western blot images and quantification of Akt activity in 3T3 cells grown on 15 kPa PDMS substrates treated with vehicle or 5 µM n-Bleb, in isotonic media, and 30 min or 120 min post-hypotonic shock. N = 3 repeats. (h) ΔpH i of HT1080 cells after treating with vehicle control, 2 µM LatA, 20 µM Y-27, and 10 µM LY29 using pHrodo Red-AM. n = 86, 109, 59, 63. (i) Normalized PIP3 level of HT1080 treated with vehicle, 10 µM LY29, and 2 µM LatA. n = 20, 14, 18. (j) Volume dynamics of HT1080 treated with 0.5 and 2 µM LatA. Hypotonic shock was applied at 30 min. All cells were treated 1 h prior to hypotonic shock. (k) NHE1 activity of 3T3 versus HT1080 predicted by the model. 3T3 parameters are set as indicated in . HT1080 parameters are the same as 3T3 excepted for and as indicated in the figure. (c-e, g-i) Error bars represent STD. (b, f, j) Error bars represent SEM. (b-e, h-i) Mann Whitney U-tests. (g) Two-way ANOVA test. (a) Scale bar = 20 µM.

    Article Snippet: PIP3 indicator pcDNA3.1 AktPH-mCherry (Addgene 67301) and membrane indicator pHR GFP-CaaX(Addgene 113020) were used to generate a ratiometric image to monitor PIP3 level.

    Techniques: Transfection, Membrane, Control, Comparison, Western Blot, Activity Assay, MANN-WHITNEY

    (a-b) Normalized PIP3 dynamics of 3T3 cells treated with 10 µM LY29 (a) and 20 µM Y-27 (b). PIP3 level is quantified by the ratio of the PIP3 indicator (AktPH-mCherry) and the membrane indicator (GFP-CaaX) at the cell boundary. (c) Representative western blot images and quantifications of Akt activity in 3T3 cells, treated with vehicle, 2 µM LatA, and 20 µM Y-27 for 30 min. (d) Representative western blot images and quantifications of Akt activity in 3T3 and HT1080 cells treated with vehicle or 40 µM EIPA. (e) Chemical module activity of 3T3 versus HT1080 predicted by the model. (f,g) NHE1 (f) and the chemical module (g) activities modeled by reducing , or both. Only by reducing both parameters was able to reproduce the insensitive response in HT1080. (h) Volume dynamics of 3T3 Piezo1 KO cells treated with 0.2 µM Trametinib (Trame) with hypotonic shock applied at 30 min. (e-g) 3T3 parameters are set as: , n 12 = n 32 = n 13 = n 23 = ™ 2, R 12 = R 23 = R 31 = 1 / 5. HT1080 parameters are the same as 3T3 excepted for and . All results are normalized to a base activity level at . (a-c, h) Error bars represent SEM. (d) Error bars represent STD.

    Journal: bioRxiv

    Article Title: Chemical Compensation to Mechanical Loss in Cell Mechanosensation

    doi: 10.1101/2025.04.02.646640

    Figure Lengend Snippet: (a-b) Normalized PIP3 dynamics of 3T3 cells treated with 10 µM LY29 (a) and 20 µM Y-27 (b). PIP3 level is quantified by the ratio of the PIP3 indicator (AktPH-mCherry) and the membrane indicator (GFP-CaaX) at the cell boundary. (c) Representative western blot images and quantifications of Akt activity in 3T3 cells, treated with vehicle, 2 µM LatA, and 20 µM Y-27 for 30 min. (d) Representative western blot images and quantifications of Akt activity in 3T3 and HT1080 cells treated with vehicle or 40 µM EIPA. (e) Chemical module activity of 3T3 versus HT1080 predicted by the model. (f,g) NHE1 (f) and the chemical module (g) activities modeled by reducing , or both. Only by reducing both parameters was able to reproduce the insensitive response in HT1080. (h) Volume dynamics of 3T3 Piezo1 KO cells treated with 0.2 µM Trametinib (Trame) with hypotonic shock applied at 30 min. (e-g) 3T3 parameters are set as: , n 12 = n 32 = n 13 = n 23 = ™ 2, R 12 = R 23 = R 31 = 1 / 5. HT1080 parameters are the same as 3T3 excepted for and . All results are normalized to a base activity level at . (a-c, h) Error bars represent SEM. (d) Error bars represent STD.

    Article Snippet: PIP3 indicator pcDNA3.1 AktPH-mCherry (Addgene 67301) and membrane indicator pHR GFP-CaaX(Addgene 113020) were used to generate a ratiometric image to monitor PIP3 level.

    Techniques: Membrane, Western Blot, Activity Assay