Journal: The Kaohsiung Journal of Medical Sciences

Article Title: MicroRNA ‐936/ ERBB4 /Akt axis exhibits anticancer properties of gastric cancer through inhibition of cell proliferation, migration, and invasion

doi: 10.1002/kjm2.12304

Figure Lengend Snippet: ERBB4 was reversely correlated with the expression of miR‐936 in gastric cancer. A, Dual luciferase reporter assay was performed to detect the binding effects of miR‐936 on 3'UTR of ERBB4. The predicted binding sequences of them were shown. ** P < .01 vs the WT‐ERBB4 3'UTR + mimic‐NC. B, RT‐qPCR and Western blot were performed to check the mRNA and protein expression levels of ERBB4 in MGC803 cells with miR‐936 overexpression. *** P < .001 vs the mimic‐NC. C, Relative mRNA and protein expression of ERBB4 were also evaluated in MKN‐45 cells after miR‐936 mimic transfection. *** P < .001 vs the mimic‐NC. D, Gastric MGC803 and MKN‐45 cells were transfected with miR‐936 inhibitor, and the expression of miR‐936 was checked by RT‐qPCR after transfection. *** P < .001 vs the inhibitor‐NC. E and F, The expression of ERBB4 was detected at both mRNA and protein levels after miR‐936 inhibitor transfection in MGC803 and MKN‐45 cells, respectively. ** P < .01, *** P < .001 vs inhibitor‐NC. ERBB4, Erb‐B2 receptor tyrosine kinase 4; MT, mutant; RT‐qPCR, reverse transcription‐quantitative polymerase chain reaction; WT, wildtype

Article Snippet: Information of primary antibody was: Akt (1:1000, Cell signaling technology, CST, USA), P‐Akt Ser473 (1:1000, CST), P‐Akt Thr308 (1:1000, CST), c‐Myc (1:2000, Proteintech, China), CyclinD1 (1:1000, Proteintech), Matrix metalloprotease 2 (MMP2) (1:1000, Proteintech), MMP9 (1:1000, Proteintech), cleaved‐caspase 3 (1:1000, Affinity, China), cleaved‐PARP (1:1000, CST), ERBB4 (1:500, Proteintech) and GAPDH (1:10000, Proteintech).

Techniques: Expressing, Luciferase, Reporter Assay, Binding Assay, Quantitative RT-PCR, Western Blot, Over Expression, Transfection, Mutagenesis, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: MicroRNA ‐936/ ERBB4 /Akt axis exhibits anticancer properties of gastric cancer through inhibition of cell proliferation, migration, and invasion

doi: 10.1002/kjm2.12304

Figure Lengend Snippet: ERBB4 overexpression reversed cell proliferation and invasion induced by miR‐936 overexpression alone in gastric cancer cells. MKN‐45 cells were cotransfected with miR‐936 mimic and ERBB4‐overexpressed plasmid for subsequent experiments. A, Cell proliferation was measured by CCK‐8 assay. B, Cell apoptosis was checked after PI/Annexin V‐FITC double staining. C, Cell migration was assessed by wound healing assay. D, Transwell assay was performed to detect cell invasion. E, Relative protein expression of ERBB4, Akt, P‐Akt Thr308 , P‐Akt Ser473 , c‐Myc, MMP2 and cleaved‐PARP was detected by western blot. * P < .05, ** P < .01, *** P < .001, ns = no statistical significance vs the mimic‐NC + vector; # P < .05, ## P < .01, ### P < .001, ns = no statistical significance vs the miR‐936 mimic + vector. ERBB4, Erb‐B2 receptor tyrosine kinase 4; Ov, overexpression; CCK‐8, cell‐counting kit‐8; PI, propidium; FITC, fluorescein isothiocyanate; MMP, matrix metallopeptidase; PARP, poly(ADP‐ribose) polymerase

Article Snippet: Information of primary antibody was: Akt (1:1000, Cell signaling technology, CST, USA), P‐Akt Ser473 (1:1000, CST), P‐Akt Thr308 (1:1000, CST), c‐Myc (1:2000, Proteintech, China), CyclinD1 (1:1000, Proteintech), Matrix metalloprotease 2 (MMP2) (1:1000, Proteintech), MMP9 (1:1000, Proteintech), cleaved‐caspase 3 (1:1000, Affinity, China), cleaved‐PARP (1:1000, CST), ERBB4 (1:500, Proteintech) and GAPDH (1:10000, Proteintech).

Techniques: Over Expression, Plasmid Preparation, CCK-8 Assay, Double Staining, Migration, Wound Healing Assay, Transwell Assay, Expressing, Western Blot, Cell Counting