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