rat somatotroph pituitary derived cell line gh3  (ATCC)

Journal: Frontiers in Endocrinology

Article Title: Potential antitumoral effects of SRPK1 inhibition through modulation of VEGF splicing in pituitary somatotroph tumoral cells

doi: 10.3389/fendo.2025.1667327

Figure Lengend Snippet: Effect of SRPIN340 and SPHINX31 on SRPK1 localization. Representative immunofluorescence images of SRPK1 in GH4C1 cells exposed to SRPIN340 or SPHINX31 (10µM) or not for 2 h and treated with EGF for 1 h. SRPK1 is represented in green whereas nuclei were counterstained in blue with DAPI. White arrows are shown to indicate subcellular localization of SRPK1 (cytoplasmatic or nuclear). The images shown are representative of three independent experiments. Scale bar: 10µm.

Article Snippet: Rat pituitary tumoral GH4C1 cells (ATCC CCL-82.2) and GH3 cells (ATCC CCL-82.1) were cultured in F10 or F12K medium (Capricorn Scientific, Ebsdorfergrund, DE), respectively, supplemented with 15% horse serum (HS), 2.5% fetal bovine serum (FBS), 2 mM glutamine and antibiotics, all supplied by Gibco (ThermoFisher Scientific, Waltham, MA, USA).

Techniques: Immunofluorescence

Journal: Frontiers in Endocrinology

Article Title: Potential antitumoral effects of SRPK1 inhibition through modulation of VEGF splicing in pituitary somatotroph tumoral cells

doi: 10.3389/fendo.2025.1667327

Figure Lengend Snippet: Effect of SRPIN340 and SPHINX31 on SRPK1 activity. Representative Western blot images and densitometrical analysis of phosphorylated SR proteins (pSRSF1, pSRSF5, pSRSF6, pSRSF4) related to their respective GAPDH loading control from GH4C1 cells treated with SRPIN340 or SPHINX31 (10µM) for 2 h and with EGF for 1 h. Cells treated with EGF only were used as control. Experiments were repeated at least three times. *p < 0.05 and **p < 0.01 vs control.

Article Snippet: Rat pituitary tumoral GH4C1 cells (ATCC CCL-82.2) and GH3 cells (ATCC CCL-82.1) were cultured in F10 or F12K medium (Capricorn Scientific, Ebsdorfergrund, DE), respectively, supplemented with 15% horse serum (HS), 2.5% fetal bovine serum (FBS), 2 mM glutamine and antibiotics, all supplied by Gibco (ThermoFisher Scientific, Waltham, MA, USA).

Techniques: Activity Assay, Western Blot, Control

Journal: Frontiers in Endocrinology

Article Title: Potential antitumoral effects of SRPK1 inhibition through modulation of VEGF splicing in pituitary somatotroph tumoral cells

doi: 10.3389/fendo.2025.1667327

Figure Lengend Snippet: SRPIN340 and SPHINX31 action on cell growth and cell apoptosis. Increasing concentration (0.1 nM – 1000 nM) of SRPIN340 or SPHINX31 were tested in GH4C1 cells in cell proliferation (A) and cell viability (C) experiments for 72h, by BrdU incorporation assay and MTT assay, respectively. (B) Cell proliferation in GH4C1 cells silenced for SRPK1 for 72 and corresponding representative immunoblotting indicating SRPK1 silencing are shown. (D) Cell apoptosis measured by caspase3/7 activity in GH4C1 cells treated with two doses of SRPIN340 or SPHINX31 (1µM-10µM) for 48 h For each set of analysis, each determination was done in triplicate and experiments were repeated at least three times. *, p < 0.05 and **, p < 0.01 vs control untreated cells or control siRNA transfected cells.

Article Snippet: Rat pituitary tumoral GH4C1 cells (ATCC CCL-82.2) and GH3 cells (ATCC CCL-82.1) were cultured in F10 or F12K medium (Capricorn Scientific, Ebsdorfergrund, DE), respectively, supplemented with 15% horse serum (HS), 2.5% fetal bovine serum (FBS), 2 mM glutamine and antibiotics, all supplied by Gibco (ThermoFisher Scientific, Waltham, MA, USA).

Techniques: Concentration Assay, BrdU Incorporation Assay, MTT Assay, Western Blot, Activity Assay, Control, Transfection

Journal: Frontiers in Endocrinology

Article Title: Potential antitumoral effects of SRPK1 inhibition through modulation of VEGF splicing in pituitary somatotroph tumoral cells

doi: 10.3389/fendo.2025.1667327

Figure Lengend Snippet: Anti-migratory effects of SRPIN340 and SPHINX31. Analysis of boyden chamber assay showing the quantification of GH4C1 cells migrated on the bottom part of the membrane after 18 h exposure to 1µM or 10µM of SRPIN340 or SPHINX31. Each determination was done in triplicate and experiments were repeated at least three times. *p < 0.05, **p < 0.01 and ***p < 0.001 vs control untreated cells.

Article Snippet: Rat pituitary tumoral GH4C1 cells (ATCC CCL-82.2) and GH3 cells (ATCC CCL-82.1) were cultured in F10 or F12K medium (Capricorn Scientific, Ebsdorfergrund, DE), respectively, supplemented with 15% horse serum (HS), 2.5% fetal bovine serum (FBS), 2 mM glutamine and antibiotics, all supplied by Gibco (ThermoFisher Scientific, Waltham, MA, USA).

Techniques: Boyden Chamber Assay, Membrane, Control

Journal: Frontiers in Endocrinology

Article Title: Potential antitumoral effects of SRPK1 inhibition through modulation of VEGF splicing in pituitary somatotroph tumoral cells

doi: 10.3389/fendo.2025.1667327

Figure Lengend Snippet: Effects of SRPIN340 and SPHINX31 on VEGF164/165a expression. (A) Analysis of vegf164a mRNA expression by RT-qPCR in GH4C1 cells treated with SRPIN340 or SPHINX31 (10 µM) for 3 h Untreated cells were used as control. Each determination was done in triplicate and experiments were repeated at least three times. **p<0.01 vs respective control. (B) VEGF164a protein levels were measured by ELISA immunoassay in culture media of GH4C1 cells incubated with SRPIN340 or SPHINX31 (10 µM) for 24 h Each determination was done in triplicate and experiments were repeated at least three times. *p < 0.05 vs control untreated cells. (C) Immunoblotting showing VEGF165a protein expression in one GH-PitNET primary culture (2#). Cells were stimulated with SRPIN340 100 nM or octreotide 10 nM for 3 h Densitometrical analysis refers to VEGF165a/GAPDH ratio and values are related to control untreated cells.

Article Snippet: Rat pituitary tumoral GH4C1 cells (ATCC CCL-82.2) and GH3 cells (ATCC CCL-82.1) were cultured in F10 or F12K medium (Capricorn Scientific, Ebsdorfergrund, DE), respectively, supplemented with 15% horse serum (HS), 2.5% fetal bovine serum (FBS), 2 mM glutamine and antibiotics, all supplied by Gibco (ThermoFisher Scientific, Waltham, MA, USA).

Techniques: Expressing, Quantitative RT-PCR, Control, Enzyme-linked Immunosorbent Assay, Incubation, Western Blot

Journal: Frontiers in Endocrinology

Article Title: Potential antitumoral effects of SRPK1 inhibition through modulation of VEGF splicing in pituitary somatotroph tumoral cells

doi: 10.3389/fendo.2025.1667327

Figure Lengend Snippet: Antitumoral effects of SRPK1 inhibitors are reverted by recombinant VEGF164a. (A) Analysis of cell migration and (B) cell viability tested in GH4C1 cells co-incubated with the recombinant rat VEGF164a protein (50 ng/ml) and SRPIN340 or SPHINX31 (1 µM) for 18 and 72 h, respectively. Each determination was done in triplicate and experiments were repeated at least three times, *p < 0.05 vs respective untreated control.

Article Snippet: Rat pituitary tumoral GH4C1 cells (ATCC CCL-82.2) and GH3 cells (ATCC CCL-82.1) were cultured in F10 or F12K medium (Capricorn Scientific, Ebsdorfergrund, DE), respectively, supplemented with 15% horse serum (HS), 2.5% fetal bovine serum (FBS), 2 mM glutamine and antibiotics, all supplied by Gibco (ThermoFisher Scientific, Waltham, MA, USA).

Techniques: Recombinant, Migration, Incubation, Control