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    MedChemExpress eipa
    (A) A schematic of the fluorescence exclusion method (FXm) and representative differential interference contrast (DIC) and epifluorescence images. (B) DIC images of HT1080 cells before and 5, 15, and 60 min after hypotonic shock. (C and D) Cell volume tracking of HT1080 (C) and MDA-MB-231 (D) cells before and after hypotonic shock. Hypotonic solutions were injected into FXm channels at 30 min. Insets in (D) show volume imaged at a 1 min frame rate for MDA-MB-231 cells ( n = 35). (E) Representative DIC images of <t>NIH</t> <t>3T3</t> cells before and 5, 15, and 60 min after hypotonic shock. (F–I) Cell volume tracking of NIH 3T3 (F), MCF-10A (G), WI-38 (H), and RPE-1 (I) cells before and after hypotonic shock. Insets in (G) and (I) show volume imaged at a 1 min frame rate for MCF-10A ( n = 29) and RPE-1 ( n = 55) cells, respectively. (C, D, and F–I) Error bars represent the standard error of the mean (SEM). Statistical tests of all time series plots can be found in – . Black arrows indicate the time of osmotic shock. (A, B, and E) Scale bars, 20 μm.
    Eipa, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 15 article reviews
    eipa - by Bioz Stars, 2026-02
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    Images

    1) Product Images from "Cytoskeletal activation of NHE1 regulates mechanosensitive cell volume adaptation and proliferation"

    Article Title: Cytoskeletal activation of NHE1 regulates mechanosensitive cell volume adaptation and proliferation

    Journal: Cell reports

    doi: 10.1016/j.celrep.2024.114992

    (A) A schematic of the fluorescence exclusion method (FXm) and representative differential interference contrast (DIC) and epifluorescence images. (B) DIC images of HT1080 cells before and 5, 15, and 60 min after hypotonic shock. (C and D) Cell volume tracking of HT1080 (C) and MDA-MB-231 (D) cells before and after hypotonic shock. Hypotonic solutions were injected into FXm channels at 30 min. Insets in (D) show volume imaged at a 1 min frame rate for MDA-MB-231 cells ( n = 35). (E) Representative DIC images of NIH 3T3 cells before and 5, 15, and 60 min after hypotonic shock. (F–I) Cell volume tracking of NIH 3T3 (F), MCF-10A (G), WI-38 (H), and RPE-1 (I) cells before and after hypotonic shock. Insets in (G) and (I) show volume imaged at a 1 min frame rate for MCF-10A ( n = 29) and RPE-1 ( n = 55) cells, respectively. (C, D, and F–I) Error bars represent the standard error of the mean (SEM). Statistical tests of all time series plots can be found in – . Black arrows indicate the time of osmotic shock. (A, B, and E) Scale bars, 20 μm.
    Figure Legend Snippet: (A) A schematic of the fluorescence exclusion method (FXm) and representative differential interference contrast (DIC) and epifluorescence images. (B) DIC images of HT1080 cells before and 5, 15, and 60 min after hypotonic shock. (C and D) Cell volume tracking of HT1080 (C) and MDA-MB-231 (D) cells before and after hypotonic shock. Hypotonic solutions were injected into FXm channels at 30 min. Insets in (D) show volume imaged at a 1 min frame rate for MDA-MB-231 cells ( n = 35). (E) Representative DIC images of NIH 3T3 cells before and 5, 15, and 60 min after hypotonic shock. (F–I) Cell volume tracking of NIH 3T3 (F), MCF-10A (G), WI-38 (H), and RPE-1 (I) cells before and after hypotonic shock. Insets in (G) and (I) show volume imaged at a 1 min frame rate for MCF-10A ( n = 29) and RPE-1 ( n = 55) cells, respectively. (C, D, and F–I) Error bars represent the standard error of the mean (SEM). Statistical tests of all time series plots can be found in – . Black arrows indicate the time of osmotic shock. (A, B, and E) Scale bars, 20 μm.

    Techniques Used: Fluorescence, Injection

    (A) Fractional cell volume change after exposure to ion transporter inhibitors: ouabain (Oua; an NKA inhibitor), EIPA (an NHE inhibitor), bumetanide (bume; an NKCC inhibitor), DCPIB (a VRAC inhibitor), and 4-AP (a VGKC inhibitor). Cell volumes were tracked for 1–4 h until fully adapted (see for details). n HT1080 = 36, 60, 50, 61, 47, 36, and 47; n 3T3 = 213, 94, 147, 76, 62, 100, and 147. (B and C) EIPA-treated HT1080 (B) and NIH 3T3 (C) cell volume dynamics under hypotonic shock. The inset in (B) shows a 1 min frame rate (vehicle, n = 53; EIPA, n = 25). (D) NIH 3T3 cell scramble control (SC) and shNHE1 volume dynamics under hypotonic shock. (E) Relative intracellular pH change ( ΔpHi ) for HT1080 and NIH 3T3 cells. Intracellular pH (pHi) is measured by pHrodo-red AM and calibrated by standard pH buffers. ΔpHi is calculated by tracking single-cell pHi. Hypotonic shock was applied at 5 min. (F and G) Oua-treated HT1080 (F) and NIH 3T3 (G) cell volume under hypotonic shock. (H and I) Representative DIC images of Oua-treated HT1080 (H) and NIH 3T3 (I) cells before and 10 and 120 min after hypotonic shock. (J and K) Computer simulation of HT1080 (J) and NIH 3T3 (K) cells with hypotonic shock and inhibition of NKA and NHE. The inset in (K) shows the NHE activation function, where α NHE is the permeability coefficient of NHE. (A) Error bars indicate standard deviation. Kruskal-Wallis tests followed by Dunn’s multiple-comparisons test were conducted between datasets and the vehicle control of the corresponding cell type. (B–G) Error bars indicate the SEM. Mann-Whitney U tests for two-condition comparisons were conducted at each time point post shock, and p values are plotted as a function of time in . (H and I) Scale bars, 20 μm.
    Figure Legend Snippet: (A) Fractional cell volume change after exposure to ion transporter inhibitors: ouabain (Oua; an NKA inhibitor), EIPA (an NHE inhibitor), bumetanide (bume; an NKCC inhibitor), DCPIB (a VRAC inhibitor), and 4-AP (a VGKC inhibitor). Cell volumes were tracked for 1–4 h until fully adapted (see for details). n HT1080 = 36, 60, 50, 61, 47, 36, and 47; n 3T3 = 213, 94, 147, 76, 62, 100, and 147. (B and C) EIPA-treated HT1080 (B) and NIH 3T3 (C) cell volume dynamics under hypotonic shock. The inset in (B) shows a 1 min frame rate (vehicle, n = 53; EIPA, n = 25). (D) NIH 3T3 cell scramble control (SC) and shNHE1 volume dynamics under hypotonic shock. (E) Relative intracellular pH change ( ΔpHi ) for HT1080 and NIH 3T3 cells. Intracellular pH (pHi) is measured by pHrodo-red AM and calibrated by standard pH buffers. ΔpHi is calculated by tracking single-cell pHi. Hypotonic shock was applied at 5 min. (F and G) Oua-treated HT1080 (F) and NIH 3T3 (G) cell volume under hypotonic shock. (H and I) Representative DIC images of Oua-treated HT1080 (H) and NIH 3T3 (I) cells before and 10 and 120 min after hypotonic shock. (J and K) Computer simulation of HT1080 (J) and NIH 3T3 (K) cells with hypotonic shock and inhibition of NKA and NHE. The inset in (K) shows the NHE activation function, where α NHE is the permeability coefficient of NHE. (A) Error bars indicate standard deviation. Kruskal-Wallis tests followed by Dunn’s multiple-comparisons test were conducted between datasets and the vehicle control of the corresponding cell type. (B–G) Error bars indicate the SEM. Mann-Whitney U tests for two-condition comparisons were conducted at each time point post shock, and p values are plotted as a function of time in . (H and I) Scale bars, 20 μm.

    Techniques Used: Control, Inhibition, Activation Assay, Permeability, Standard Deviation, MANN-WHITNEY

    (A–C) Confocal images of NIH 3T3 (A), HT1080 (B), and LatA-treated NIH 3T3 cells (C) stained with the live-cell F-actin probe SPY650-FastAct before and after hypotonic shock. (D) Representative images showing elevated membrane tension measured by Flipper-TR lifetime during hypotonic shock in NIH 3T3 cells grown on glass. (E) Membrane tension of NIH 3T3 cells grown on glass and on 15 kPa PDMS substrates before and after 3, 30, or 2 h of hypotonic shock. n = 61, 41, 72, and 81 for glass and 25, 18, 27, and 40 for 15 kPa substrates. (F–H) Volume dynamics of LatA-treated NIH 3T3 (F), LatA-treated HT1080 (G), and LY29-treated NIH 3T3 (H) cells under hypotonic shock. (I) Volume dynamics of NIH 3T3 cells grown on 3, 15, and 130 kPa PDMS substrates and on glass under hypotonic shock. (J) ΔpHi for NIH 3T3 cells grown on 15 kPa substrates versus on glass. Hypotonic shock was applied at 5 min. (K) ΔpHi for LatA-treated NIH 3T3 cells versus vehicle control under hypotonic shock. (F–K) Error bars indicate SEM. Mann-Whitney U tests for two-condition comparisons were conducted at each time point post shock, and p values are plotted as a function of time in . (E) Error bars indicate standard deviation. Kruskal-Wallis tests followed by Dunn’s multiple-comparisons test were used. (A–D) Scale bars, 20 μm.
    Figure Legend Snippet: (A–C) Confocal images of NIH 3T3 (A), HT1080 (B), and LatA-treated NIH 3T3 cells (C) stained with the live-cell F-actin probe SPY650-FastAct before and after hypotonic shock. (D) Representative images showing elevated membrane tension measured by Flipper-TR lifetime during hypotonic shock in NIH 3T3 cells grown on glass. (E) Membrane tension of NIH 3T3 cells grown on glass and on 15 kPa PDMS substrates before and after 3, 30, or 2 h of hypotonic shock. n = 61, 41, 72, and 81 for glass and 25, 18, 27, and 40 for 15 kPa substrates. (F–H) Volume dynamics of LatA-treated NIH 3T3 (F), LatA-treated HT1080 (G), and LY29-treated NIH 3T3 (H) cells under hypotonic shock. (I) Volume dynamics of NIH 3T3 cells grown on 3, 15, and 130 kPa PDMS substrates and on glass under hypotonic shock. (J) ΔpHi for NIH 3T3 cells grown on 15 kPa substrates versus on glass. Hypotonic shock was applied at 5 min. (K) ΔpHi for LatA-treated NIH 3T3 cells versus vehicle control under hypotonic shock. (F–K) Error bars indicate SEM. Mann-Whitney U tests for two-condition comparisons were conducted at each time point post shock, and p values are plotted as a function of time in . (E) Error bars indicate standard deviation. Kruskal-Wallis tests followed by Dunn’s multiple-comparisons test were used. (A–D) Scale bars, 20 μm.

    Techniques Used: Staining, Membrane, Control, MANN-WHITNEY, Standard Deviation

    (A) Representative NIH 3T3 and HT1080 cell GCaMP6s snapshots before and after hypotonic shock. (B) Cytosolic Ca 2 + concentration of HT1080 cells grown on glass, NIH 3T3 cells grown on glass, and NIH 3T3 cells grown on 15 kPa PDMS substrates, measured by GCaMP6s intensity under hypotonic shock (applied at 10 min) and imaged at a 2 min frame rate. (C) NIH 3T3 single-cell GCaMP6s dynamics in isotonic medium (top) and 20 min after hypotonic shock (bottom) on glass, imaged at a 10 s frame rate. (D) The number of Ca 2 + spikes in isotonic medium versus 20–40 min under hypotonic shock for each cell type and substrate; defined as at least 2× intensity increase over baseline. n = 26, 20, 18, 31, 21, and 20. Mann-Whitney U tests. (E–G) Immunostaining of Piezo1 at the basal plane for all cell types and conditions in (D). (H and I) Volume and ΔpHi dynamics in NIH 3T3 WT cells and Piezo1 KO cells during hypotonic shock. (J) Co-staining of NHE1 and actin and their distribution along the line. (K) NSC-treated NIH 3T3 cell volume dynamics under hypotonic shock. (L–N) ΔpHi of NIH 3T3 versus HT1080 (L), NSC-treated NIH 3T3 (M), and NIH 3T3 Piezo1 KO (N) cells under 20% uniaxial mechanical stretch (applied at 5 min). (B, H, I, and K–N) Error bars indicate SEM. Mann-Whitney U tests were used to compare two conditions at each time point after osmotic shock or mechanical stretch, with p values plotted in . (D) Error bars indicate standard deviation. (A, E–G, and J) Scale bars, 20 μm.
    Figure Legend Snippet: (A) Representative NIH 3T3 and HT1080 cell GCaMP6s snapshots before and after hypotonic shock. (B) Cytosolic Ca 2 + concentration of HT1080 cells grown on glass, NIH 3T3 cells grown on glass, and NIH 3T3 cells grown on 15 kPa PDMS substrates, measured by GCaMP6s intensity under hypotonic shock (applied at 10 min) and imaged at a 2 min frame rate. (C) NIH 3T3 single-cell GCaMP6s dynamics in isotonic medium (top) and 20 min after hypotonic shock (bottom) on glass, imaged at a 10 s frame rate. (D) The number of Ca 2 + spikes in isotonic medium versus 20–40 min under hypotonic shock for each cell type and substrate; defined as at least 2× intensity increase over baseline. n = 26, 20, 18, 31, 21, and 20. Mann-Whitney U tests. (E–G) Immunostaining of Piezo1 at the basal plane for all cell types and conditions in (D). (H and I) Volume and ΔpHi dynamics in NIH 3T3 WT cells and Piezo1 KO cells during hypotonic shock. (J) Co-staining of NHE1 and actin and their distribution along the line. (K) NSC-treated NIH 3T3 cell volume dynamics under hypotonic shock. (L–N) ΔpHi of NIH 3T3 versus HT1080 (L), NSC-treated NIH 3T3 (M), and NIH 3T3 Piezo1 KO (N) cells under 20% uniaxial mechanical stretch (applied at 5 min). (B, H, I, and K–N) Error bars indicate SEM. Mann-Whitney U tests were used to compare two conditions at each time point after osmotic shock or mechanical stretch, with p values plotted in . (D) Error bars indicate standard deviation. (A, E–G, and J) Scale bars, 20 μm.

    Techniques Used: Concentration Assay, MANN-WHITNEY, Immunostaining, Staining, Standard Deviation

    (A) Representative images of N2FXm and 3D reconstructed cell and cytoplasm volumes. Nucleus volume is calculated by subtracting cytoplasm from total cell volume. (B) Normalized nucleus volume of NIH 3T3 and HT1080 cells following hypotonic shock applied at 5 min. (C) Normalized nucleus volume of NSC-treated NIH 3T3 cells and vehicle control following hypotonic shock applied at 5 min. (D) Principal-component analysis (PCA) for NIH 3T3 cells before and after hypotonic shock and with or without LatA treatment. N = 3 biological repeats. (E and F) Volcano plots showing DEGs at 2-h hypotonic shock versus in isotonic medium for NIH 3T3 (E) and HT1080 (F) cells. (G) Top 6 GSEA terms in the ChEA transcription factor database for genes with hypermethylated (top) and hypomethylated (bottom) DEG-DMRs and top 30 contributing genes (right) at 2-h hypotonic shock versus in isotonic medium for NIH 3T3 cells. (B and C) Error bars indicate SEM. Mann-Whitney U tests were used to compare two conditions at each time point after osmotic shock, and the p values are plotted in . (A) Scale bars, 10 μm.
    Figure Legend Snippet: (A) Representative images of N2FXm and 3D reconstructed cell and cytoplasm volumes. Nucleus volume is calculated by subtracting cytoplasm from total cell volume. (B) Normalized nucleus volume of NIH 3T3 and HT1080 cells following hypotonic shock applied at 5 min. (C) Normalized nucleus volume of NSC-treated NIH 3T3 cells and vehicle control following hypotonic shock applied at 5 min. (D) Principal-component analysis (PCA) for NIH 3T3 cells before and after hypotonic shock and with or without LatA treatment. N = 3 biological repeats. (E and F) Volcano plots showing DEGs at 2-h hypotonic shock versus in isotonic medium for NIH 3T3 (E) and HT1080 (F) cells. (G) Top 6 GSEA terms in the ChEA transcription factor database for genes with hypermethylated (top) and hypomethylated (bottom) DEG-DMRs and top 30 contributing genes (right) at 2-h hypotonic shock versus in isotonic medium for NIH 3T3 cells. (B and C) Error bars indicate SEM. Mann-Whitney U tests were used to compare two conditions at each time point after osmotic shock, and the p values are plotted in . (A) Scale bars, 10 μm.

    Techniques Used: Control, MANN-WHITNEY

    (A) Volcano plots showing DEGs under 2-h hypotonic shock between LatA-treated NIH 3T3 cells and vehicle control. (B) Top 4 upstream pathways identified by IPA in NIH 3T3 cells under 2-h hypotonic shock with and without LatA treatment. Red/green indicate increased/decreased measurements, and orange/blue indicate predicated activation/deactivation. (C) Gene heatmap showing expression of MAPK/ERK pathway genes identified in (B). (D) Western blot for phospho-ERK and total ERK of NIH 3T3 WT, NIH 3T3 Piezo1 KO, and HT1080 cells, with quantification of phospho-ERK/total ERK ratio. N = 4 biological replicates. Student’s t test. (E) Images and quantification of Ki-67-active cells under isotonic conditions and after 6 h of hypotonic shock with and without Trame treatment in NIH 3T3 WT, NIH 3T3 Piezo1 KO, and HT1080 cells. N = 4 biological replicates, and >200 cells were examined in each repeat. (F) Top 6 GSEA terms in the ChEA transcription factor database for genes with hypermethylated (top) and hypomethylated (bottom) DEG-DMRs at 2-h hypotonic shock with and without LatA treatment in NIH 3T3 cells. (G) Schematic of the proposed cytoskeletal activation of the NHE1 pathway. (D and E) Error bars indicate standard deviation. Mann-Whitney U tests. (E) Scale bars, 20 μm.
    Figure Legend Snippet: (A) Volcano plots showing DEGs under 2-h hypotonic shock between LatA-treated NIH 3T3 cells and vehicle control. (B) Top 4 upstream pathways identified by IPA in NIH 3T3 cells under 2-h hypotonic shock with and without LatA treatment. Red/green indicate increased/decreased measurements, and orange/blue indicate predicated activation/deactivation. (C) Gene heatmap showing expression of MAPK/ERK pathway genes identified in (B). (D) Western blot for phospho-ERK and total ERK of NIH 3T3 WT, NIH 3T3 Piezo1 KO, and HT1080 cells, with quantification of phospho-ERK/total ERK ratio. N = 4 biological replicates. Student’s t test. (E) Images and quantification of Ki-67-active cells under isotonic conditions and after 6 h of hypotonic shock with and without Trame treatment in NIH 3T3 WT, NIH 3T3 Piezo1 KO, and HT1080 cells. N = 4 biological replicates, and >200 cells were examined in each repeat. (F) Top 6 GSEA terms in the ChEA transcription factor database for genes with hypermethylated (top) and hypomethylated (bottom) DEG-DMRs at 2-h hypotonic shock with and without LatA treatment in NIH 3T3 cells. (G) Schematic of the proposed cytoskeletal activation of the NHE1 pathway. (D and E) Error bars indicate standard deviation. Mann-Whitney U tests. (E) Scale bars, 20 μm.

    Techniques Used: Control, Activation Assay, Expressing, Western Blot, Standard Deviation, MANN-WHITNEY

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Transfection, Sequencing, shRNA, Control, Knock-Out, Software



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