Fig 2: FOXC1 binds to the Srsf1 promoter to regulate Srsf1 expression.A Relative expression of Foxc1 detected by qPCR (n = 4). B, C Expression of FOXC1 was detected by western blot (n = 3). D CHIP-qPCR was utilized to detect the binding sites of FOXC1 and Srsf1 promoter region. E Relative expression of Foxc1 by qPCR (n = 4). F, G Western Blot was utilized to detect the expression of FOXC1 (n = 3). H Relative expression of Foxc1 by qPCR (n = 5). I, J The expression level of FOXC1 was detected by western blot (n = 3). K Relative expression of Srsf1 by qPCR (n = 4). L, M Western blot was utilized to detect the expression of SRSF1 (n = 3). N Relative expression of Srsf1 detected by qPCR (n = 4). O, P Western blot was utilized to detect the expression level of SRSF1 (n = 3). In (A, C, D, E, G, H, J, K, M, N, P), the data are expressed as mean ± SD, where statistical significance was calculated using Student’s t test in (A, C, D) and using one-way ANOVA with Tukey post hoc test in (F, G, I, J, L, M, O). *P < 0.05, **P < 0.01, ***P < 0.001, nsP > 0.05.

Fig 3: Effect of Srsf1 overexpression on OB apoptosis induced by GC.A qPCR to verify Srsf1 overexpression efficiency (n = 4), GV492: lentiviral vector, OE-Srsf1: overexpression of Srsf1. B, C The overexpression efficiency of SRSF1 was detected by western blot (n = 3). D, E TUNEL/DAPI staining detection of OBs TUNEL* cell number (n = 3). F–H Expression levels of cleaved-CASP3 and cleaved-CASP9 were detected by western blot (n = 3). I, J Apoptosis detected by Annexin V-FITC/PI staining (n = 4). In (A, C, D, G, H, and J), the data were expressed as mean ± SD, and statistical significance was calculated using one-way ANOVA with Tukey post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, nsP > 0.05.

Fig 4: Overview of the involvement of Srsf1 in OB apoptosis induced by GC.GC induces the upregulation of Hdac4 and decreases the level of H3K9ac and H3K27ac in the Foxc1 promoter region to inhibit the expression of FOXC1, thereby reducing the expression of Srsf1. SRSF1 can bind to the 7807–8006 nucleotide sites of Bmf pre-mRNA to regulate the alternative splicing of Bmf. Overexpression of Srsf1 reduced the expression of Bmf V2 and Bmf V3 variants, thereby inhibiting GC-induced OB apoptosis; conversely, knockdown of Srsf1 increased the expression of Bmf V2 and V3 variants, thereby promoting GC-induced OB apoptosis.

Fig 5: Effects of Srsf1 knockdown on GC-induced OB apoptosis.A qPCR to verify the knockdown efficiency of Srsf1 (n = 4), CV279: the control lentivirus vector, KD-Srsf1: knockdown of Srsf1. B, C Western Blot detection of SRSF1 knockdown efficiency (n = 4). D, E TUNEL/DAPI staining detection of OBs TUNEL* cell number (n = 3). F–H Expression levels of cleaved-CASP3 and cleaved-CASP9 were detected by western blot (n = 3). I, J Apoptosis detected by Annexin V-FITC/PI staining (n = 4). In (A, C, D, G, H, and J), the data were expressed as mean ± SD, and statistical significance was calculated using one-way ANOVA with Tukey post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, nsP > 0.05.