apc 052  (Alomone Labs)

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A, B) TRPM7 current densities and (B) TRPM7 I/V relationship of Jurkat T cells during whole-cell patch clamp experiment with Mg 2+ -free intracellular solution. TRPM7 WT (WT, gray) and KO2 Jurkat clone (KO2, orange), n (WT) = 9; n (KO2) = 10. (C) Cell counts and (D) viability of natively proliferating TRPM7 WT and KO2 Jurkat clone in RPMI medium with 10% FBS, with and without supplementation of 6 mM MgCl 2 , n = 3, measured in duplicates. (E) Cellular Mg 2+ contents quantified by ICP-MS. TRPM7 WT and KO2 Jurkat clone, cultured in regular (WT-)media without or with 6 mM MgCl 2 supplementation for 18 h ahead of sampling, n = 4. (F) Fura-2-based imaging of cytosolic Ca 2+ concentration of Jurkat T cells. Passive store release was induced with 5 μM thapsigargin at indicated time point (arrow). TRPM7 WT (WT, gray) and KO2 (KO2, orange) Jurkat clone, n (WT) = 111; n (KO2) = 59. (G) Quantification of the area under the curve (AUC) of respective curves shown in (F). (H) Representative immuno-fluorescent images of NFATc1 localization in TRPM7 WT and KO2 clone before (basal) and after 30 min stimulation (stim.) with 5 μM thapsigargin, scale bar = 2 μm. NFATc1 in red, DAPI in blue. (I, J) Representative intensity profiles of subcellular NFATc1 localization (NFATc1 in red, DAPI in blue) of Jurkat TRPM7 WT (WT, gray) and KO2 (KO2, orange) in basal state (I) and upon 30 min passive store depletion induced with 5 μM thapsigargin (J). (K) Quantification of nuclear NFATc1 levels (corresponding to AF647 signal intensity) upon stimulation of TRPM7 WT (WT, gray) and KO (KO2, orange) clone, n (WT) = 261; n (KO2) = 149. (L) Histograms and (M) quantification of up-regulated CD69 expression of Jurkat TRPM7 WT (WT, gray) and KO2 (KO2, orange) cells after overnight α-CD3 stimulation, n = 4–6. (B, F, G, H, J) Statistics: One-way ANOVA (B), Two-way ANOVA (F, G), or t test (H, J). * P < 0.05; **** P < 0.0001, n.s., not significant. Data are mean ± SD.

Article Snippet: The following antibodies were used: α-human CD4-VioBlue (REA623; Miltenyi), α-human CD45RA-APC-Vio770 (REA562; Miltenyi), α-human CD69-APC (REA824; Miltenyi), α-human CD25-VioBright515 (REA570; Miltenyi).

Techniques: Patch Clamp, Cell Culture, Sampling, Imaging, Concentration Assay, Expressing

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A, B) TRPM7 current densities and (B) TRPM7 I/V relationship of Jurkat T cells during whole-cell patch clamp experiment with Mg 2+ -free intracellular solution. Control (Ctrl, gray) and cells treated with 1 μM Apamin (Apamin, blue), n (Ctrl) = 9; n (Apamin) = 6. (C) Cell counts and (D) viability of natively proliferating Jurkat T cells in RPMI medium with 10% FBS, treated with 1 μM Apamin (Apamin, blue) or control (Ctrl, gray), n = 4. (E) Cell counts and (F) viability of natively proliferating Jurkat TRPM7 WT and KO cells in RPMI medium with 10% FBS, treated with 30 μM NS8593, 30 μM NS8593 with additional 6 mM MgCl 2 or untreated controls, n = 3, measured in duplicates. (G) Fura-2 based imaging of cytosolic Ca 2+ concentrations of Jurkat TRPM7 WT and KO cells treated with 30 μM NS8593 or left untreated. Passive store depletion was induced with 5 μM thapsigargin at indicated time point (arrow). (H) Quantification of area under the curve (AUC) of traces shown in (G), n = 24–70. (I) Representative FACS plots and gating strategy for CD69 visualization, shown for Jurkat WT cells. (J) Histogram and (K) quantification of up-regulated CD69 expression of Jurkat TRPM7 WT cells treated with 1 μM Apamin (Apamin, blue) compared with untreated controls (Ctrl, light gray), n = 3. (L) Histogram and (M) quantification of up-regulated CD69 expression of Jurkat TRPM7 WT or KO cells treated with 30 μM NS8593 or left untreated, n = 4–6. (H, K, M) Statistics: One-way ANOVA (H, M) and t test (K). ** P < 0.005, *** P < 0.001, **** P < 0.0001, n.s., not significant. Data are mean ± SD.

Article Snippet: The following antibodies were used: α-human CD4-VioBlue (REA623; Miltenyi), α-human CD45RA-APC-Vio770 (REA562; Miltenyi), α-human CD69-APC (REA824; Miltenyi), α-human CD25-VioBright515 (REA570; Miltenyi).

Techniques: Patch Clamp, Control, Imaging, Expressing

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A) Fura-2 based imaging of cytosolic Ca 2+ concentration of Jurkat T cells. Passive store release was induced with 5 μM thapsigargin at the indicated time point (arrow) of WT (black) and TRPM7 KO (red) Jurkat T cells, n (WT) = 111; n (KO) = 113. (B) Quantification of the area under the curve (AUC) of respective curves shown in (A). (C) Representative immune-fluorescence images of the NFATc1 localization in TRPM7 WT and KO cells before (basal) and after 30 min stimulation (stim.) with 5 μM thapsigargin, scale bar = 2 μm. NFATc1 in red, DAPI in blue. (D, E) Representative intensity profiles of subcellular NFATc1 localization (NFATc1 in red, DAPI in blue) of Jurkat TRPM7 WT (black) and KO (red) cells in basal state (D) and upon 30 min passive store depletion induced with 5 μM thapsigargin (E). (F) Quantification of nuclear NFATc1 levels (corresponding to AF647 signal intensity) upon stimulation of TRPM7 WT (black) and KO (red) cells, n (WT) = 261; n (KO) = 279. (G) Relative IL-2 mRNA expression levels of Jurkat TRPM7 WT (black) and KO (red) cells, n = 4. (H) Histograms and (I) quantification of up-regulated CD69 expression of Jurkat TRPM7 WT (black) and KO (red) cells after overnight stimulation with α-CD3, n = 4–6. (J) Quantification of Ca 2+ signals of TRPM7 WT Jurkat T cells, treated with 30 μM NS8593 (NS, red) or DMSO control (Ctrl, black). Passive store release was induced with 5 μM thapsigargin at indicated time point (arrow), n (Ctrl) = 95; n (NS) = 94. (K) Quantification of the area under the curve (AUC) of respective Ca 2+ signals shown in (G). (L) Representative immune-fluorescence images of NFATc1 localization of cells treated with 30 μM NS8593 (NS, red) or DMSO control (Ctrl, black) before and after 30 min stimulation with 5 μM thapsigargin, scale bar = 2 μm. (M, N) Representative intensity profiles of subcellular NFATc1 localization (NFATc1 in red, DAPI in blue) of Jurkat TRPM7 WT (black) and KO (red) cells in basal state (M) and upon 30 min passive store depletion induced with 5 μM thapsigargin (N). (O) Quantification of nuclear NFATc1 levels upon stimulation of cells treated with 30 μM NS8593 (NS, red) or DMSO control (Ctrl, black), n (Ctrl) = 196; n (NS) = 195. (P) Relative IL- 2 mRNA expression levels of cells treated with 30 μM NS8593 (NS, red) or DMSO control (Ctrl, black), n = 7. (Q) Histograms and (R) quantification of up-regulated CD69 expression of cells treated with 30 μM NS8593 (NS, red) or DMSO control (Ctrl, black) after α-CD3 stimulation, n = 6–7. (B, D, E, F, H, J, K, M) Statistics: t test (B, D, F, H, J, M) and Mann-Whitney U test (E, K). ** P < 0.005; *** P < 0.0005; **** P < 0.0001 and n.s., not significant. Data are mean ± SD.

Article Snippet: The following antibodies were used: α-human CD4-VioBlue (REA623; Miltenyi), α-human CD45RA-APC-Vio770 (REA562; Miltenyi), α-human CD69-APC (REA824; Miltenyi), α-human CD25-VioBright515 (REA570; Miltenyi).

Techniques: Imaging, Concentration Assay, Fluorescence, Expressing, Control, MANN-WHITNEY

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A, B) Representative FACS plots and gating strategy to confirm identity of isolated naïve CD4 T cells and (B) total CD4 T cells. (C) Representative traces of Fura-2-based imaging of cytosolic Ca 2+ concentrations following anti-CD3/CD28 stimulation in CD4 T cells. Antibodies bound to microscopy chamber bottom with cells sinking down in saline containing 2 mM Ca 2+ during running measurement, coming to rest in focus plane with contact to stimulation antibodies. Cells treated with 1 μM Apamin (Apamin, blue) or control (Ctrl, gray). (C, D, E) Quantification of area under the curve (D) and oscillation frequency (E) of data shown in (C), n = 29–37. (F) Representative FACS plots and gating strategy for CD69 and CD25 in total CD4 T cells. (G, H) Quantification of flow cytometry data of NS8593 dose-dependent up-regulation of CD69 (G) and CD25 (H) expression on total CD4 T cells, 48 h after anti-CD3/CD28 stimulation or PMA/ionomycin stimulation, respectively, n = 3–4. (I, J) Quantification of flow cytometry data of up-regulation of CD69 (I) and CD25 (J) expression on total CD4 T cells treated with 1 μM Apamin (Apamin, blue) or control (Ctrl, gray), 48 h after anti-CD3/CD28 stimulation or PMA/ionomycin stimulation, respectively, n = 3. (K) Respective FACS plots and gating strategy for CD4 T cell proliferation, shown for control cells. (L) Respective quantification of NS8593 dose-dependent proliferation of total CD4 T cells, with and without supplementation of 6 mM MgCl 2 , corresponding to , n = 4–7. (M) Representative histograms of dose-dependent proliferation (CSFE dye dilution) of total CD4 T cells in presence 1 μM Apamin in comparison to control, with (right) and without (left) supplementation of 6 mM MgCl 2 . Cells gated on T cell population, single cells and CD4 + T cells. Color code as in (N). (N) Respective quantification of proliferation of total CD4 T cells treated with 1 μM Apamin (Apamin, blue) or control (Ctrl, gray), with and without supplementation of 6 mM MgCl 2 , n = 5–8. (O) Respective quantification of Waixenicin A dose-dependent proliferation of total CD4 T cells, with and without supplementation of 6 mM MgCl 2 , corresponding to , n = 4–7. (D, E, G, H, I, J, L, N, O) Statistics: t test (D, E, I, J) and one-way ANOVA (G, H, L, N, O). * P < 0.05; ** P < 0.005, **** P < 0.0001 and n.s., not significant. Data are mean ± SD.

Article Snippet: The following antibodies were used: α-human CD4-VioBlue (REA623; Miltenyi), α-human CD45RA-APC-Vio770 (REA562; Miltenyi), α-human CD69-APC (REA824; Miltenyi), α-human CD25-VioBright515 (REA570; Miltenyi).

Techniques: Isolation, Imaging, Microscopy, Saline, Control, Flow Cytometry, Expressing, Comparison

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A) IL-2 quantification in supernatant of naïve CD4 T cells 48 h after α-CD3/α-CD28 stimulation, n = 4–5. (B, C, D, E) Histograms and quantification of up-regulated activation markers CD69 (B, C) and CD25 (D, E) in naïve CD4 T lymphocytes 48 h after stimulation. Cells were treated with 30 μM NS8593 or DMSO control, both with (Ctrl, blue; NS, orange) and without (Ctrl, black; NS, red) supplementation of 6 mM MgCl 2 . (F) IL-2 quantification in supernatant of total CD4 T cells 48 h after α-CD3/α-CD28 stimulation or cells treated with 30 μM NS8593 or DMSO control, both with (Ctrl, blue; NS, orange) and without (Ctrl, black; NS, red) supplementation of 6 mM MgCl 2 , n = 4–5. (G, H, I, J) Histograms and quantification of up-regulated activation markers CD69 (G, H) and CD25 (I, J) in total CD4 T lymphocytes 48 h after stimulation. Cells treated with either 30 μM NS8593 or DMSO control, both with (Ctrl, blue; NS, orange) and without (Ctrl, black; NS, red) supplementation of 6 mM MgCl 2 . (K) Representative TRPM7 I/V relationships of total CD4 T cells obtained via whole-cell patch clamp with Mg 2+ -free intracellular solution. Cells were treated with 10 μM Waixenicin A (WxA, green) or EtOH control (Ctrl, black). (L, M, N, O) Histograms and quantification of up-regulated activation markers CD69 (L, M) and CD25 (N, O) in total CD4 T lymphocytes 48 h after stimulation. Cells treated with 10 μM Waixenicin A or EtOH control, both with (Ctrl, blue; WxA, light green) and without (Ctrl, black; WxA, green) supplementation of 6 mM MgCl 2 , n = 7. (P) Representative histograms of dose-dependent proliferation (CSFE dye dilution) of total CD4 T cells in presence of various NS8593 concentrations, with (right) and without (left) supplementation of 6 mM MgCl 2 . Cells gated on T cell population, single cells and CD4 + T cells. Color code as in (Q). (Q) Respective quantification of NS8593 dose-dependent proliferation of total CD4 T cells, with and without supplementation of 6 mM MgCl 2 , corresponding to (P), n = 4–7. (R) Representative histograms of dose-dependent proliferation (CSFE dye dilution) of total CD4 T cells in presence of various Waixenicin A concentrations, with (right) and without (left) supplementation of 6 mM MgCl 2 . Cells gated on T cell population, single cells and CD4 + T cells. Color code as in (S). (S) Respective quantification of Waixenicin A dose-dependent proliferation of total CD4 T cells, with and without supplementation of 6 mM MgCl 2 , corresponding to (S), n = 4–8. (A, C, E, F, H, J, M, O, Q, S) Statistics: one-way ANOVA (A, C, E, F, H, J, M, O, Q, S). * P < 0.05; ** P < 0.005; *** P < 0.0005; **** P < 0.0001 and n.s., not significant. Data are mean ± SD.

Article Snippet: The following antibodies were used: α-human CD4-VioBlue (REA623; Miltenyi), α-human CD45RA-APC-Vio770 (REA562; Miltenyi), α-human CD69-APC (REA824; Miltenyi), α-human CD25-VioBright515 (REA570; Miltenyi).

Techniques: Activation Assay, Control, Patch Clamp

Journal: Life Science Alliance

Article Title: TRPM7 and magnesium orchestrate human CD4 T-cell activation and differentiation

doi: 10.26508/lsa.202503357

Figure Lengend Snippet: (A) Schematic description of naïve CD4 T-cell differentiation towards FOXP3-expressing regulatory T cells and RORƔt-expressing T H 17 cells, including respective cytokine polarization milieus. (B, C) Percentages of CD45RA − cells and (C) CD25 + CD127 lo cells upon polarization of naïve CD4 T cells toward iT reg cells in various NS8593 concentrations (red) compared with DMSO control (Ctrl, black), n = 6–7. (D, E) Representative FACS histograms and (E) quantification of FOXP3 expression levels of CD25 + CD127 lo iT reg cells upon 6 d polarization of naïve CD4 T cells in presence of various NS8593 concentrations (red) or DMSO control (Ctrl, black), n = 6–7. (F, G) Percentages of CD45RA − cells and (G) CD25 + CD127 lo cells upon polarization of naïve CD4 T cells toward iT reg cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 7. (H, I) Representative FACS histograms and (I) quantification of FOXP3 expression levels of CD25 + CD127 lo iT reg cells upon 6 d polarization of naïve CD4 T cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 6. (J) Percentages of CCR6 + cells upon polarization of naïve CD4 T cells towards iT H 17 cells in presence of various NS8593 concentrations (red) compared with DMSO control (Ctrl, black), n = 6. (K, L) Representative FACS histograms and (L) quantification of RORƔt expression levels of CCR6 + iT H 17 cells upon 6 d polarization of naïve CD4 T cells in presence of various NS8593 concentrations (red) or DMSO control (Ctrl, black), n = 4–6. (M) Percentages of CCR6 + cells upon polarization of naïve CD4 T cells towards iT H 17 cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 6. (N, O) Representative FACS histograms and (O) quantification of RORƔt expression levels of CCR6 + iT H 17 cells upon 6 d polarization of naïve CD4 T cells in presence of 6 mM MgCl 2 (MgCl 2 , blue) compared with H 2 O control (Ctrl, black), n = 5. (P) Graphical summary of TRPM7-(in)dependent T-cell activation and differentiation towards iT reg and iT H 17 cells. Pharmacological blockade of TRPM7 reduces intracellular Mg 2+ levels, leads to reduced Ca 2+ signaling and results in reduced IL-2 secretion, impaired up-regulation of T-cell activation markers CD69 and CD25, and diminished proliferation upon TCR stimulus (left). TRPM7 inhibition during polarization of naïve CD4 T cells into iT reg cells preserves FOXP3 + signals of CD25 + CD127 lo iT reg cells. Polarization of naïve CD4 T cells into iT H 17 cells results in augmented RORƔt expression in the presence of 6 mM Mg 2+ , which is reduced upon TRPM7 inhibition, highlighting the need for Mg 2+ uptake and related TRPM7-dependent intracellular signaling for iT H 17 cell polarization (right). (B, C, E, F, G, I, J, L, M, O) Statistics: one-way ANOVA (B, C, E, J, L) and t test (F, G, I, M, O). * P < 0.05; ** P < 0.005; *** P < 0.0005; **** P < 0.0001 and n.s., not significant. Data are mean ± SD.

Article Snippet: The following antibodies were used: α-human CD4-VioBlue (REA623; Miltenyi), α-human CD45RA-APC-Vio770 (REA562; Miltenyi), α-human CD69-APC (REA824; Miltenyi), α-human CD25-VioBright515 (REA570; Miltenyi).

Techniques: Cell Differentiation, Expressing, Control, Activation Assay, Inhibition

Journal: bioRxiv

Article Title: Differential contribution of HCN1 and HCN4 to the synchronisation of sinoatrial pacemaker cells

doi: 10.64898/2025.12.16.694405

Figure Lengend Snippet: The SAN network forms the intrinsic leading pacemaker region of the heart driven by If-conducting cells. (A) Preparation of a gelatine-inflated heart indicating the SAN region (dashed line). (B) Fluorescence imaging of an anti-HCN1 (red signal) and anti-HCN4 (green signal) immunostained whole-mount right atrial preparation of a WT heart, revealing the anatomical extension (head, body, tail region) and spatial HCN1/HCN4 channel expression profile of the SAN. (C) Isolated SAN pacemaker cell (elongated cell) during a patch clamp recording. (D) Family of current traces (If) recorded from a WT pacemaker cell of the sinoatrial node using the voltage protocol shown on the top. From a holding potential of −50 mV, a series of voltage pulses ranging from −140 to - 20 mV (duration 5 s, delta V: 15 mV) were applied. (E) Mean activation curve of native If recorded from WT SAN pacemaker cells (n=13). Additional abbreviations: CT, crista terminalis; SVC, superior vena cava.

Article Snippet: After permeabilization (0.5% Triton X100, 20% DMSO in PBS) and blocking in 5% NDS (Normal Donkey Serum), the tissue was incubated with guinea-pig polyclonal HCN1 antibody (1:500; Alomone Labs, Jerusalem, Israel) and rabbit polyclonal HCN4 antibody (1:500; Alomone Labs).

Techniques: Fluorescence, Imaging, Expressing, Isolation, Patch Clamp, Activation Assay

Journal: Frontiers in Neuroanatomy

Article Title: Transmitter and ion channel profiles of saccadic omnipause neurons and cholinergic non-omnipause neurons in human nucleus raphe interpositus

doi: 10.3389/fnana.2025.1670220

Figure Lengend Snippet: Immunoperoxidase labeling of HCN subunits in human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained for HCN1 (left) and HCN2 (right). HCN1 (left) is labeled only in OPNs, which are ensheathed by perineuronal nets (red arrows) and lacks completely in cholinergic non-OPNs (green arrows) (middle). On the other hand, HCN2 (right) immunolabeling is found in both neuron groups. (B) HCN4 immunolabeling (right) is ubiquitously found both in OPNs (red arrows) in cholinergic non-OPNs (green arrows). Scale bar represents 50 μm.

Article Snippet: HCN4 subunit was detected with polyclonal guinea pig antibody (Cat #: APC-052-GP (formerly AGP-004); RRID: AB_2340957; Alomone Labs, Jerusalem, Israel), which recognizes an intracellular epitope at the N-terminus, corresponding to amino acid residues 119–155 of human HCN4.

Techniques: Labeling, Staining, Immunolabeling

Journal: Frontiers in Neuroanatomy

Article Title: Transmitter and ion channel profiles of saccadic omnipause neurons and cholinergic non-omnipause neurons in human nucleus raphe interpositus

doi: 10.3389/fnana.2025.1670220

Figure Lengend Snippet: Immunoperoxidase labeling of HCN subunits in human nucleus raphe interpositus (RIP). (A) Consecutive 5 μm thick coronal paraffin sections through RIP stained for HCN1 (left) and HCN2 (right). HCN1 (left) is labeled only in OPNs, which are ensheathed by perineuronal nets (red arrows) and lacks completely in cholinergic non-OPNs (green arrows) (middle). On the other hand, HCN2 (right) immunolabeling is found in both neuron groups. (B) HCN4 immunolabeling (right) is ubiquitously found both in OPNs (red arrows) in cholinergic non-OPNs (green arrows). Scale bar represents 50 μm.

Article Snippet: HCN4 , Guinea Pig/Polyclonal , Hyperpolarization-activated cyclic nucleotide-gated channel 4 , Alomone Labs Jerusalem BioPark (JBP) , AB_2340957 , 1:200.

Techniques: Labeling, Staining, Immunolabeling

Journal: The Journal of Physiological Sciences : JPS

Article Title: Doxorubicin-induced sinus node dysfunction associated with mitochondria and nuclear impairment in a mouse model

doi: 10.1016/j.jphyss.2025.100047

Figure Lengend Snippet: Doxorubicin-induced cardiomyopathy modelling in mice. A. A work flow of the modelling of doxorubicin (DXB)-induce cardiomyopathy in C57BL/6 N adult male mice. Saline was injected as a vehicle (Veh) control. B. The time course of the body weight in Veh- and DXB-treated mice. While the body weight of Veh mice incrementally gained, that of DXB mice decreased. n = 6 in Veh and 10 in DXB. C. Heart weight-to-tibial length ratio in Veh- and DXB-treated mice. n = 6 in Veh and 5 in DXB. D and E. Picrosirius red-stained sections (D) and averaged fibrosis area (E) of the left ventricular myocardium in Veh- and DXB-treated mice. n = 4/group. Scale bars = 20 μm. F. Representative picrosirius red-stained sections of the SN region in Veh- or DXB-treated mice and enlarged area labelled with HCN4, a pacemaker channel as a SN marker, indicated by rectangles in picrosirius red images. Scale bars = 100 μm. G. Averaged fibrosis area of the SN region in Veh- and DXB-treated mice. n = 4/group. * p < 0.05 vs Veh, determined by unpaired t -test.

Article Snippet: After blocking with 0.1 % bovine serum albumin in PBS, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-HCN4 antibodies (1:100, APC-052, Alomone Labs).

Techniques: Saline, Injection, Control, Staining, Marker

Journal: The Journal of Physiological Sciences : JPS

Article Title: Doxorubicin-induced sinus node dysfunction associated with mitochondria and nuclear impairment in a mouse model

doi: 10.1016/j.jphyss.2025.100047

Figure Lengend Snippet: Chronic effect of doxorubicin (DXB) on expression levels of genes responsible for pacemaking. A and B. Gene expression levels of HCN4 pacemaker channel and Ca 2+ regulators in the sinus node (A) and the right atrial myocardium (B) in vehicle (Veh)- and DXB-treated mice. n = 6 in SN, n = 3 in right atrium, * p < 0.05 vs Veh determined by unpaired t -test.

Article Snippet: After blocking with 0.1 % bovine serum albumin in PBS, the sections were incubated overnight at 4 °C with rabbit polyclonal anti-HCN4 antibodies (1:100, APC-052, Alomone Labs).

Techniques: Expressing, Gene Expression