anti cav1 3  (Alomone Labs)

Journal: Scientific Reports

Article Title: Voltage-gated calcium channels as key regulators of neuronal differentiation in the immortalized dorsal root ganglion neuronal cell line F11

doi: 10.1038/s41598-026-44595-1

Figure Lengend Snippet: Differentiation of F11 cells increased the expression of Ca V 1.2 and Ca V 1.3 VGCCs. Expression of Ca V 1.2 VGCCs in a the cell membrane (non-permeabilized cells) and b the whole cell (permeabilized cells) of non-differentiated (–) and differentiated F11 cells (+) (mean ± S.D.). c – f Representative images of non-differentiated and differentiated F11 cells stained for nuclei (blue) and Ca V 1.2 VGCCs (orange) under non-permeabilized and permeabilized conditions. Expression of Ca V 1.3 VGCCs in g the cell membrane (non-permeabilized cells) and h the whole cell (permeabilized cells) of non-differentiated (–) and differentiated F11 cells (+) (mean ± S.D.). i – l Representative images of non-differentiated and differentiated F11 cells stained for nuclei (blue) and Ca V 1.3 VGCCs (orange) under non-permeabilized and permeabilized conditions. Data and images for both channels are representative of one experiment out of three independent experiments ( N = 3). Each data point represents one replicate within the experiment ( n = 4). Values in each graph are expressed as a percentage relative to the expression of Ca V 1.2 or Ca V 1.3 VGCCs, respectively, in non-differentiated F11 cells in each condition, which is defined as 100%. *** p < 0.001, ** p < 0.01; Welch’s t test. Scale bar in microphotographs = 100 μm.

Article Snippet: Cells were exposed to a solution containing anti-Ca V 1.2 channel (ACC-003; Alomone Labs) and anti-Ca V 1.3 channel rabbit antibodies (ACC-005; Alomone Labs), each diluted 1:500 in DMEM at 4 °C.

Techniques: Expressing, Membrane, Staining

Journal: Scientific Reports

Article Title: Voltage-gated calcium channels as key regulators of neuronal differentiation in the immortalized dorsal root ganglion neuronal cell line F11

doi: 10.1038/s41598-026-44595-1

Figure Lengend Snippet: Ca V 1.3 overexpression enhanced neurite outgrowth under basal conditions, while Ca V 1.2 and Ca V 1.3 increased the 30 mM KCl-evoked intracellular Ca²⁺ signal. a Effect of 30 mM KCl on intracellular calcium signal in F11 cells after overexpression (Ovr.) of Ca V 1.2 and Ca V 1.3 channels compared to control F11 cells (mean ± S.D.). Data are representative of one experiment out of three independent experiments ( N = 3). Each data point represents one replicate within the experiment ( n = 9). *** p < 0.001, * p < 0.05; ANOVA followed by Dunnett’s post-hoc analysis. Effect of the overexpression of Ca V 1.2 and Ca V 1.3 under basal conditions in b , c not permeabilized and d , e permeabilized cells. Data are representative of one experiment out of three independent experiments ( N = 3). Each data point represents one replicate within the experiment ( n = 4). *** p < 0.001, ** p < 0.01; Welch’s t test. f – m Representative images of each condition with nuclei stained in blue and channels stained in orange. n Effect of overexpression of Ca V 1.2 and Ca V 1.3 channels on maximum neurite length compared to control F11 cells (mean ± S.D). Data are representative of one experiment out of three independent experiments ( N = 3). Each data point represents one replicate within the experiment ( n = 6). * p < 0.05; ANOVA followed by Dunnett’s post-hoc analysis. Representative images of o control F11 cells and F11 cells after overexpression of p Ca V 1.2 channels and q Ca V 1.3 channels stained for nuclei (red) and β-tubulin (green). r Effect of overexpression of Ca V 1.2 and Ca V 1.3 channels on CellROX Green intensity, representing intracellular ROS concentrations, compared to control F11 cells (mean ± S.D). Data are representative of one experiment out of four independent experiments ( N = 4). Each data point represents one replicate within the experiment ( n = 6). Representative images of s control F11 cells and F11 cells after overexpression of t Ca V 1.2 and u Ca V 1.3 channels stained for nuclei (blue) and with CellROX Green. Values in graphs are expressed as a percentage relative to the effect observed in control F11 cells transfected with an empty plasmid (100%). Scale bar in microphotographs = 100 μm.

Article Snippet: Cells were exposed to a solution containing anti-Ca V 1.2 channel (ACC-003; Alomone Labs) and anti-Ca V 1.3 channel rabbit antibodies (ACC-005; Alomone Labs), each diluted 1:500 in DMEM at 4 °C.

Techniques: Over Expression, Control, Staining, Transfection, Plasmid Preparation

Journal: Scientific Reports

Article Title: Voltage-gated calcium channels as key regulators of neuronal differentiation in the immortalized dorsal root ganglion neuronal cell line F11

doi: 10.1038/s41598-026-44595-1

Figure Lengend Snippet: Ca V 1.2 and Ca V 1.3 overexpression under differentiation conditions impaired acquisition of neuronal features and increased ROS. a Effect of 30 mM KCl on intracellular calcium signal in F11 cells after overexpression (Ovr.) of Ca V 1.2 and Ca V 1.3 channels compared to control F11 cells (mean ± S.D.). Data are representative of one experiment out of three independent experiments ( N = 3). Each data point represents one replicate within the experiment ( n = 9). *** p < 0.001; ANOVA followed by Dunnett’s post-hoc analysis. Effect of the overexpression of Ca V 1.2 and Ca V 1.3 under differentiation conditions in b , c not permeabilized and d , e permeabilized cells. Data are representative of one experiment out of three independent experiments ( N = 3). Each data point represents one replicate within the experiment ( n = 4). ** p < 0.01, * p < 0.05; Welch’s t test. f – m Representative images of each condition with nuclei stained in blue and channels stained in orange. n Effect of overexpression of Ca V 1.2 and Ca V 1.3 channels on maximum neurite length compared to control F11 cells (mean ± S.D). Data are representative of one experiment out of three independent experiments ( N = 3). Each data point represents one replicate within the experiment ( n = 6). *** p < 0.001, * p < 0.05; ANOVA followed by Dunnett’s post-hoc analysis. Representative images of o control F11 cells and F11 cells after overexpression of p Ca V 1.2 channels and q Ca V 1.3 channels stained for nuclei (red) and β-tubulin (green). r Effect of overexpression of Ca V 1.2 and Ca V 1.3 channels on CellROX Green intensity, representing intracellular ROS concentrations, compared to control F11 cells (mean ± S.D). Data are representative of one experiment out of four independent experiments ( N = 4). Each data point represents one replicate within the experiment ( n = 6). *** p < 0.001, * p < 0.05; ANOVA followed by Dunnett’s post-hoc analysis. Representative images of ( s ) control F11 cells and F11 cells after overexpression of t Ca V 1.2 and u Ca V 1.3 channels stained for nuclei (blue) and with CellROX Green. Values in graphs are expressed as a percentage relative to the effect observed in control F11 cells transfected with an empty plasmid (100%). Scale bar in microphotographs = 100 μm.

Article Snippet: Cells were exposed to a solution containing anti-Ca V 1.2 channel (ACC-003; Alomone Labs) and anti-Ca V 1.3 channel rabbit antibodies (ACC-005; Alomone Labs), each diluted 1:500 in DMEM at 4 °C.

Techniques: Over Expression, Control, Staining, Transfection, Plasmid Preparation

Journal: Science Advances

Article Title: α-Synuclein expression is required for somatodendritic dopamine release and immediate early gene induction

doi: 10.1126/sciadv.ady6978

Figure Lengend Snippet: ( A ) Representative single-channel currents recorded in a cell-attached configuration from cultured WT and SKO SN DA neurons in the presence of N-, P/Q-, and R-type channel blockers show unitary currents (downward deflections) due to LTCC openings elicited by a voltage ramp (bottom trace). ( B ) Single-channel current amplitude was identical for LTCC in WT and SKO mice (ns by two-way ANOVA; n = 8 WT and 9 SKO). ( C ) Ensemble average P O - V relationships show no change in the probability of channel openings between WT and SKO neurons (ns by two-way ANOVA). ( D ) Population data confirm no change in maximal open channel probability (ns by t test). ( E and G ) Representative confocal images of cultured hippocampal neurons (14 days postplating) from WT and SKO mice immunostained for α1C (E) or α1D (G) subunits of the LTCC, as well as αSyn and pan-neuronal microtubule-associated protein 2 (MAP2). Note that staining intensity of the α1D subunit was low, making the quantification of the signal less reliable. Scale bar, 10 μm. ( F and H ) Analysis of average cytosolic and membrane α1C (F) and α1D (H) staining intensity ( n = 70 to 78 cells in each group from three independent experiments. * P < 0.05, ** P < 0.01, or **** P < 0.0001 by t test).

Article Snippet: Primary antibodies include anti-actin (mouse monoclonal, Sigma-Aldrich, #A5441, RRID: AB_476744; 1:1000), anti–glyceraldehyde-3-phosphate dehydrogenase (mouse monoclonal, Proteintech, #60004-1-Ig, RRID: AB_2107436; 1:1000), anti–pCREB (Ser 133 , rabbit monoclonal, Cell Signaling Technology, #9198S; 1:500), anti-CREB1 (rabbit polyclonal, ABclonal, #A11064, RRID: AB_2758389; 1:500), anti–c-Fos (rabbit monoclonal, Cell Signaling Technology, #2250S, RRID: AB_2247211; 1:1000), anti-Ca v 1.2 α1C (rabbit polyclonal, Proteintech, #21774-1-AP, RRID: AB_2878918; 1:500), and anti-Ca v 1.3 α1D (rabbit polyclonal, Alomone Labs, #ACC-005, RRID: AB_2039775; 1:100).

Techniques: Cell Culture, Staining, Membrane