Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: Primer sequences used for qRT-PCR

Article Snippet: Next, the samples were electroblotted onto PVDF membranes and blocked in 5% skim milk for 2 h. Thereafter, the membranes were kept overnight with primary antibodies against GAPDH (ab181602; Abcam), proliferating cell nuclear antigen (PCNA; ab18197; Abcam, Cambridge, MA, USA), cleaved-caspase 3 (ab2302; Abcam), E-cadherin (ab231303; Abcam), N-cadherin (ab207608; Abcam), vimentin (ab137321; Abcam) or PAX6 (ab195045; Abcam) and secondary antibody (ab205718; Abcam) for 2 h. The bands were exposed using an ECL kit (Beyotime).

Techniques:

Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: miR-132-3p directly interacted with PAX6. (a) The complementary sequences between PAX6 and miR-132-3p. (b) The interaction between miR-132-3p and PAX6 was demonstrated by dual-luciferase reporter assay. (c) The protein level of PAX6 in Y79 and WERI-Rb-1 cells transfected with miR-NC or miR-132-3p was measured by western blot assay. (d) The expression of miR-132-3p in Y79 and WERI-Rb-1 cells transfected with anti-NC or anti-miR-132-3p was determined by qRT-PCR assay. (e) The protein level of PAX7 in anti-NC or anti-miR-132-3p transfected Y79 and WERI-Rb-1 cells was measured with western blot assay. (f) The protein level of PAX6 in Y79 and WERI-Rb-1 cells with sh-NC + anti-NC, sh-circRNF20#1 + anti-NC or sh-circRNF20#1 + anti-miR-132-3p was measured through western blot assay. (g and h) The protein level of PAX6 in RB tissues and cells was examined via western blot assay. *** P < 0.001.

Article Snippet: Next, the samples were electroblotted onto PVDF membranes and blocked in 5% skim milk for 2 h. Thereafter, the membranes were kept overnight with primary antibodies against GAPDH (ab181602; Abcam), proliferating cell nuclear antigen (PCNA; ab18197; Abcam, Cambridge, MA, USA), cleaved-caspase 3 (ab2302; Abcam), E-cadherin (ab231303; Abcam), N-cadherin (ab207608; Abcam), vimentin (ab137321; Abcam) or PAX6 (ab195045; Abcam) and secondary antibody (ab205718; Abcam) for 2 h. The bands were exposed using an ECL kit (Beyotime).

Techniques: Luciferase, Reporter Assay, Transfection, Western Blot, Expressing, Quantitative RT-PCR

Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: circRNF20 regulated RB cell malignant behaviors by miR-132-3p and PAX6. Sh-NC, sh-circRNF20#1, sh-circRNF20#1 + anti-NC, sh-circRNF20#1 + anti-miR-132-3p, sh-circRNF20#1 + vector or sh-circRNF20#1 + PAX6 was transfected into Y79 and WERI-Rb-1 cells. (a) The protein level of PAX6 in Y79 and WERI-Rb-1 cells was examined by qRT-PCR assay. (b–d) The viability, proliferation and colony formation of Y79 and WERI-Rb-1 cells were tested by CCK-8 assay, EDU assay and colony formation assay. (e) The apoptosis of Y79 and WERI-Rb-1 cells was analyzed by flow cytometry analysis. (f and g) The migration and invasion of Y79 and WERI-Rb-1 cells were assessed by wound-healing assay and transwell assay. (h) The protein levels of PCNA, cleaved-caspase 3, E-cadherin, N-cadherin and vimentin in Y79 and WERI-Rb-1 cells were measured by western blot assay. *** P < 0.001.

Article Snippet: Next, the samples were electroblotted onto PVDF membranes and blocked in 5% skim milk for 2 h. Thereafter, the membranes were kept overnight with primary antibodies against GAPDH (ab181602; Abcam), proliferating cell nuclear antigen (PCNA; ab18197; Abcam, Cambridge, MA, USA), cleaved-caspase 3 (ab2302; Abcam), E-cadherin (ab231303; Abcam), N-cadherin (ab207608; Abcam), vimentin (ab137321; Abcam) or PAX6 (ab195045; Abcam) and secondary antibody (ab205718; Abcam) for 2 h. The bands were exposed using an ECL kit (Beyotime).

Techniques: Plasmid Preparation, Transfection, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Colony Assay, Flow Cytometry, Migration, Wound Healing Assay, Transwell Assay, Western Blot

Journal: Open Medicine

Article Title: circRNF20 aggravates the malignancy of retinoblastoma depending on the regulation of miR-132-3p/PAX6 axis

doi: 10.1515/med-2022-0483

Figure Lengend Snippet: circRNF20 silencing blocked tumor growth in vivo . (a and b) The volume and weight of xenograft tumor were examined. (c) The levels of ki-67 and PAX6 in xenograft tumors were examined by IHC assay. (d) The levels of circRNF20 and miR-132-3p in xenograft tumors were detected by qRT-PCR assay. (e) The protein levels of PCNA, cleaved-caspase 3, PAX6, E-cadherin, N-cadherin and vimentin in xenograft tumors were measured via western blot assay. ** P < 0. 01, *** P < 0.001.

Article Snippet: Next, the samples were electroblotted onto PVDF membranes and blocked in 5% skim milk for 2 h. Thereafter, the membranes were kept overnight with primary antibodies against GAPDH (ab181602; Abcam), proliferating cell nuclear antigen (PCNA; ab18197; Abcam, Cambridge, MA, USA), cleaved-caspase 3 (ab2302; Abcam), E-cadherin (ab231303; Abcam), N-cadherin (ab207608; Abcam), vimentin (ab137321; Abcam) or PAX6 (ab195045; Abcam) and secondary antibody (ab205718; Abcam) for 2 h. The bands were exposed using an ECL kit (Beyotime).

Techniques: In Vivo, Quantitative RT-PCR, Western Blot

Journal: Stem cell research

Article Title: Generation and characterization of induced pluripotent stem cells from breast cancer patients carrying ATM mutations

doi: 10.1016/j.scr.2023.103246

Figure Lengend Snippet: Reagents details.

Article Snippet: , Rabbit Anti-Pax6 , 1:200 , Thermo Fisher Scientific Cat #42–6600 , RRID: AB_2533534.

Techniques: Immunocytochemistry, Virus, Plasmid Preparation