Fig 1: Silencing of FENDRR inhibits the tumorigenesis of gastric cancer cells in vitro via inactivation of PI3K/Akt signaling. (A) The protein expression levels of MST1R, caspase-3, p-Akt, Akt, p-ERK and ERK in Kato III cells were determined by western blot. (B) The relative protein expression of MST1R was quantified by normalizing to ß-actin. (C) The ratio of cleaved caspase-3/procaspase-3 was calculated. (D) The ratio of p-Akt/Akt was calculated. (E) The ratio of p-ERK/ERK was calculated. FENDRR, FOXF1 adjacent non-coding developmental regulatory RNA; MST1R, macrophage stimulating 1 receptor; OE, overexpression vector; NC, non-coding control; siRNA, short interfering RNA; ERK, extracellular signal related kinase; p, phosphorylated **P<0.01 vs. NC; ##P<0.01 vs. siRNA-FENDRR.
Fig 2: SRSF1 binds to MST1R. (A) Equal amounts of total lysates were applied to co-immunoprecipitation with IgG, anti-SRSF1 or anti-MST1R magnetic beads. Immunoprecipitation analysis of Kato III cells transfected with NC or siRNA- FENDRR . (B) The enrichment of SRSF1 was detected by RNA pull-down. FENDRR, FOXF1 adjacent non-coding developmental regulatory RNA; SRSF1, serine/arginine rich splicing factor 1; MST1R, macrophage stimulating 1 receptor; NC, non-coding control; siRNA, short interfering RNA; IgG, immunoglobulin G; ctrl, control. **P<0.01 vs. probe-ctrl.
Fig 3: MST1R and RON ?160 are upregulated in Kato III cells. (A) The protein expression levels of MST1R in GES-1, Kato III and MKN-45 cells were determined by western blot. The relative expression was quantified by normalizing to ß-actin. (B) The expression levels of RON ?160 in GES-1, Kato III and MKN-45 cells were investigated by immunofluorescence staining. Green fluorescence indicates RON ?160. Blue fluorescence indicates DAPI. (C) Electrophoresis of MST1R, RON ?160 and ß-actin polymerase chain reaction products on the agarose gel showed a unique band with expected sizes for each gene for GES-1, Kato III and MKN-45 cells. DNA expression was quantified by normalizing to ß-actin. MST1R, macrophage stimulating 1 receptor; RON ?160, recepteur d'origine nantais. **P<0.01 vs. GES-1 cells.
Fig 4: Silencing of FENDRR suppresses the expression of MST1R and altered the distribution of RON ?160 in Kato III cells. (A) Kato III cells were transfected with NC or FENDRR siRNA for 24 h. The efficiency of transfection was measured by reverse transcription-quantitative PCR. (B) The protein expression levels of MST1R and SRSF1 in Kato III cells were determined by western blot. The relative protein expression levels of MST1R and SRSF1 were quantified by normalizing to ß-actin. (C) The expression level of RON ?160 in Kato III cells was determined by immunofluorescence staining. (D) Electrophoresis of MST1R, RON ?160 and ß-actin polymerase chain reaction products on the agarose gel showed a unique band with expected sizes for each gene for Kato III cells. DNA expression was quantified by normalizing to ß-actin. FENDRR, FOXF1 adjacent non-coding developmental regulatory RNA; SRSF1, serine/arginine rich splicing factor 1; MST1R, macrophage stimulating 1 receptor; RON ?160, recepteur d'origine nantais; NC, non-coding control; siRNA, short interfering RNA. **P<0.01 vs. NC.
Fig 5: Knockdown of SRSF1 significantly inactivates MST1R and RON ?160 in Kato III cells. (A) Kato III cells were transfected with NC, siRNA-SRSF1-1, siRNA-SRSF1-2 or siRNA-SRSF1-3 for 24 h and then western blotting was used to detect the efficiency of transfection. (B) The relative protein expression of SRSF1 was quantified by normalizing to ß-actin. (C) The expression of RON ?160 in Kato III cells was measured by immunofluorescence staining. (D) The protein expression level of MST1R in Kato III cells was determined by western blot. The relative expression levels of MST1R were quantified by normalizing to ß-actin. (E) Electrophoresis of MST1R, RON ?160 and ß-actin polymerase chain reaction products on the agarose gel showed a unique band with expected sizes for each gene for Kato III cells. DNA expression was quantified by normalizing to ß-actin. SRSF1, serine/arginine rich splicing factor 1; MST1R, macrophage stimulating 1 receptor; RON ?160, recepteur d'origine nantais; NC, non-coding control; siRNA, short interfering RNA. **P<0.01 vs. control.
Supplier Page from Abcam for Anti-RON (phospho Y1353) antibody [EPR5598]