Journal: Bioscience Reports

Article Title: Modulating Galectin-1 in human osteoblast-like cells alters mineralisation and influences the expression of genes associated with osteoblasts and osteocytes

doi: 10.1042/BSR20253890

Figure Lengend Snippet: ( A ) Endogenous LGALS1 expression in unmodified Saos2 cells cultured under basal (light grey) and osteogenic (dark grey) conditions. Overexpression ( B ) and knockdown ( C ) of LGALS1 at the gene level under basal (i) and osteogenic (ii) culture conditions. EV = empty vector, LGALS1 = LGALS1 overexpressing cells, NT = non-target control, L-423 = LGALS1 knockdown. The error bars represent the standard deviation from three to five experimental replicates. * indicates P <0.05 (Student’s unpaired t -test, with Welsh’s correction applied to all except panel (Ci)).

Article Snippet: Saos2 cells were seeded onto a 6-well plate in basal media lacking penicillin/streptomycin and cultured until 70%–80% confluent and then transfected with 2.5 μg of pCMV6-AC-GFP human LGALS1 per well (RG204674, NM_002305 ; Origene, U.K.) using Lipofectamine 3000 (L3000) (Invitrogen) according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Over Expression, Knockdown, Plasmid Preparation, Control, Standard Deviation

Journal: Bioscience Reports

Article Title: Modulating Galectin-1 in human osteoblast-like cells alters mineralisation and influences the expression of genes associated with osteoblasts and osteocytes

doi: 10.1042/BSR20253890

Figure Lengend Snippet: Cell viability is shown as RFU following ( A ) LGALS1 over-expression (EV = empty vector control, LGALS1 = LGALS1 overexpression) and ( B ) LGALS1 knockdown (NT = non-target control, L423 = shRNA to LGALS1 ). Error bars represent standard deviation from seven technical replicates. * indicates P <0.05 (Student’s unpaired t -test, with Welsh’s correction applied in panel (A)).

Article Snippet: Saos2 cells were seeded onto a 6-well plate in basal media lacking penicillin/streptomycin and cultured until 70%–80% confluent and then transfected with 2.5 μg of pCMV6-AC-GFP human LGALS1 per well (RG204674, NM_002305 ; Origene, U.K.) using Lipofectamine 3000 (L3000) (Invitrogen) according to the manufacturer’s instructions.

Techniques: Over Expression, Plasmid Preparation, Control, Knockdown, shRNA, Standard Deviation

Journal: Bioscience Reports

Article Title: Modulating Galectin-1 in human osteoblast-like cells alters mineralisation and influences the expression of genes associated with osteoblasts and osteocytes

doi: 10.1042/BSR20253890

Figure Lengend Snippet: ( A ) Phalloidin staining (red) in empty vector control cells, LGALS1 overexpressing cells, non-target control cells, and LGALS1 knock-down cells (L-423). Scale bars = 100 μm. Images representative of three experimental replicates. ( B ) Quantification of cell area and circularity showing the mean ± standard deviation. No statistical differences were observed (one-way ANOVA).

Article Snippet: Saos2 cells were seeded onto a 6-well plate in basal media lacking penicillin/streptomycin and cultured until 70%–80% confluent and then transfected with 2.5 μg of pCMV6-AC-GFP human LGALS1 per well (RG204674, NM_002305 ; Origene, U.K.) using Lipofectamine 3000 (L3000) (Invitrogen) according to the manufacturer’s instructions.

Techniques: Staining, Plasmid Preparation, Control, Knockdown, Standard Deviation

Journal: Bioscience Reports

Article Title: Modulating Galectin-1 in human osteoblast-like cells alters mineralisation and influences the expression of genes associated with osteoblasts and osteocytes

doi: 10.1042/BSR20253890

Figure Lengend Snippet: Panels ( A i) and ( A ii) represent basal conditions, and panels ( B i) and ( B ii) represent osteogenic conditions. Blue bars represent the non-target control (NT), and pink bars represent L-423 ( LGALS1 knockdown). Expression is shown relative to the ACTB housekeeping gene. Error bars represent the standard deviation from three to five experimental replicates. * indicates P <0.05 (unpaired Student’s t -test with or without Welsh’s correction, or where the expression was not normally distributed, a Mann–Whitney U test was performed). Heatmaps show the mean expression levels of osteogenic marker genes using Z -score normalisation. Blue indicates high expression, and white indicates low expression. The mean of three to five experimental replicates was used for these comparisons.

Article Snippet: Saos2 cells were seeded onto a 6-well plate in basal media lacking penicillin/streptomycin and cultured until 70%–80% confluent and then transfected with 2.5 μg of pCMV6-AC-GFP human LGALS1 per well (RG204674, NM_002305 ; Origene, U.K.) using Lipofectamine 3000 (L3000) (Invitrogen) according to the manufacturer’s instructions.

Techniques: Control, Knockdown, Expressing, Standard Deviation, MANN-WHITNEY, Marker

Journal: Bioscience Reports

Article Title: Modulating Galectin-1 in human osteoblast-like cells alters mineralisation and influences the expression of genes associated with osteoblasts and osteocytes

doi: 10.1042/BSR20253890

Figure Lengend Snippet: Panels ( A i) and ( A ii) represent basal conditions, and panels ( B i) and ( B ii) represent osteogenic conditions. Blue bars represent the empty vector control (EV), and pink bars represent LGALS1 ( LGALS1 overexpression). Expression is shown relative to the ACTB housekeeping gene. Error bars represent the standard deviation from three to five experimental replicates. * indicates P <0.05 (unpaired Student’s t -test with or without Welsh’s correction, or where the expression was not normally distributed, a Mann–Whitney U test was performed). Heatmaps show the mean expression levels of osteogenic marker genes using Z -score normalisation. Blue indicates high expression, and white indicates low expression. The mean of three to five experimental replicates was used for these comparisons.

Article Snippet: Saos2 cells were seeded onto a 6-well plate in basal media lacking penicillin/streptomycin and cultured until 70%–80% confluent and then transfected with 2.5 μg of pCMV6-AC-GFP human LGALS1 per well (RG204674, NM_002305 ; Origene, U.K.) using Lipofectamine 3000 (L3000) (Invitrogen) according to the manufacturer’s instructions.

Techniques: Plasmid Preparation, Control, Over Expression, Expressing, Standard Deviation, MANN-WHITNEY, Marker

Journal: Bioscience Reports

Article Title: Modulating Galectin-1 in human osteoblast-like cells alters mineralisation and influences the expression of genes associated with osteoblasts and osteocytes

doi: 10.1042/BSR20253890

Figure Lengend Snippet: ( A ) ALP activity measured in conditioned media taken at day 0 (basal culture) and after 7, 14, and 21 days in osteogenic media (EV = empty vector control, LGALS1 = LGALS1 overexpressing cells). No significant differences were observed (mixed effects model with Bonferroni correction). ( B ) Alizarin red staining for calcium deposits after 21 days of osteogenic culture shown pre- and post-staining in (i) empty vector control cells and (ii) cells overexpressing LGALS1 . (iii) Quantification of alizarin red staining showing the percentage area that is positively stained. * indicates P <0.05 (Student’s unpaired t -test with Welsh’s correction) ( C ) Picrosirius red staining for collagen deposition after 21 days of osteogenic culture in (i) empty vector control cells and (ii) cells overexpressing LGALS1 . (iii) Quantification of Picrosirius red staining showing the percentage area that is positively stained. Error bars represent standard deviation from three experimental replicates. No significant differences were observed (Student’s unpaired t -test). Scale bars = 500 μm.

Article Snippet: Saos2 cells were seeded onto a 6-well plate in basal media lacking penicillin/streptomycin and cultured until 70%–80% confluent and then transfected with 2.5 μg of pCMV6-AC-GFP human LGALS1 per well (RG204674, NM_002305 ; Origene, U.K.) using Lipofectamine 3000 (L3000) (Invitrogen) according to the manufacturer’s instructions.

Techniques: Activity Assay, Plasmid Preparation, Control, Staining, Standard Deviation

Journal: Bioscience Reports

Article Title: Modulating Galectin-1 in human osteoblast-like cells alters mineralisation and influences the expression of genes associated with osteoblasts and osteocytes

doi: 10.1042/BSR20253890

Figure Lengend Snippet: ( A ) ALP activity measured in conditioned media taken at day 0 (basal culture) and after 7, 14, and 21 days in osteogenic media (NT = non-target control, L-423 = LGALS1 knockdown cells). *indicates P <0.05 (mixed effects model with Bonferroni correction). ( B ) Staining of mineralised calcium phosphate crystals (green) after 21 days of osteogenic culture in (i) NT control, (ii) LGALS1 knockdown cells. Scale bars = 1000 μm and (iii) quantification staining of mineralised calcium phosphate crystals showing the percentage area that is positively stained. * P <0.05 (unpaired student’s t -test). ( C ) Picrosirius staining for collagen after 21 days of osteogenic culture in (i) NT control, (ii) LGALS1 knockout cells. Scale bars = 500 μm and (iii) quantification of staining showing the percentage area that is positively stained. Error bars represent standard deviation from three experimental replicates. No significant difference is observed (Mann–Whitney U test).

Article Snippet: Saos2 cells were seeded onto a 6-well plate in basal media lacking penicillin/streptomycin and cultured until 70%–80% confluent and then transfected with 2.5 μg of pCMV6-AC-GFP human LGALS1 per well (RG204674, NM_002305 ; Origene, U.K.) using Lipofectamine 3000 (L3000) (Invitrogen) according to the manufacturer’s instructions.

Techniques: Activity Assay, Control, Knockdown, Staining, Knock-Out, Standard Deviation, MANN-WHITNEY

Journal: Cancers

Article Title: Targeting Galectin-1 with Triptolide Induces Ferroptosis in Oral Squamous Cell Carcinoma

doi: 10.3390/cancers18050782

Figure Lengend Snippet: Galectin-1 regulates the expression of ferroptosis-associated genes in OSCC cells. ( A ) SAS cells were transfected with LGALS1-targeting siRNA (si-Gal-1) or scrambled control siRNA. After 48 h, mRNA expression levels of LGALS1, GPX4, SLC7A11, and FTH1 were measured using quantitative PCR (qPCR). ( B ) SAS cells were transfected with a pCMV-Galectin-1 overexpression plasmid. Following 48 h, the expression of LGALS1, GPX4, SLC7A11, and FTH1 was quantified by qPCR. Gene expression values were normalized to GAPDH and calculated using the 2 −ΔΔCt method. Data represent mean ± SD from three independent biological replicates ( n = 3). * p < 0.05 compared with control.

Article Snippet: For overexpression, cells were transfected with a pCMV-Galectin-1 plasmid (OriGene, Rockville, MD, USA) using jetPRIME ® transfection reagent (Polyplus-transfection, Illkirch, France) following the manufacturer’s protocol.

Techniques: Expressing, Transfection, Control, Real-time Polymerase Chain Reaction, Over Expression, Plasmid Preparation, Gene Expression