- PA5-61288 - Invitrogen Antibodies MEIS3 antibody

Cai P, Xie Y, Dong M, Zhu Q
BioMed research international 2020;2020:2046248

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of MEIS3 in Lane 1: Marker (kDa) 250, 130, 95, 72, 55, 36, 28, 17, 10; Lane 2: Human cell line RT-4. Samples were probed using a MEIS3 Polyclonal Antibody (Product # PA5-61288).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of MEIS3 in Lane 1: Marker (kDa) 250, 130, 95, 72, 55, 36, 28, 17, 10; Lane 2: Human cell line RT-4. Samples were probed using a MEIS3 Polyclonal Antibody (Product # PA5-61288).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of MEIS3 in human cell line MCF7 shows positivity in nucleus but excluded from the nucleoli. Samples were probed using a MEIS3 Polyclonal Antibody (Product # PA5-61288).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of MEIS3 in human small intestine shows strong cytoplasmic end membranous positivity in glandular cells. Samples were probed using a MEIS3 Polyclonal Antibody (Product # PA5-61288).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 (a) Based on the IC50 of cetuximab in 43 colon cancer cell lines from the Genomics of Drug Sensitivity in Cancer (GDSC) dataset, we selected SNU-C1, CL-34, and SW1417 as cetuximab-resistant cell lines and HT-155, HCC2998, and DiFi-09844799147 as cetuximab-sensitive cell lines. (b) The RNA expression profile in both cetuximab-sensitive and cetuximab-resistant cells was obtained, and genes with low expression in cetuximab-resistant cell lines were identified. (c) Pipeline of transfection of sgRNA library and drug selection. (d) NGS readout was used to quantify sgRNA. (e) mRNA from bulk tumor tissue was extracted from 17 cetuximab-sensitive and nine cetuximab-resistant patients, and expression of MEIS3 was identified by qPCR. (f) Bulk tumor tissue protein from eight cetuximab-sensitive and six cetuximab-resistant cell lines was extracted, and the expression of MEIS3 was detected by western blot ( n = 3). (g) The relationship between the expression of MEIS3 and the overall survival rate of COAD patients in TCGA database.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 (a) mRNA was extracted from 11 colon cancer cell lines, and the expression of MEIS3 was detected by western blot ( n = 3). (b) The LOVO cells were transfected with random shRNA, CON vector, and MEIS3 shRNA, and the knockdown efficiency was confirmed by western blot after 48 h. (c) The LOVO-CON and MEIS3 shRNA cells were treated with different concentrations of cetuximab for 48 h, and PI staining was used to detect changes in the cell cycle. (d) The LOVO-CON and MEIS3 shRNA cells were treated with different concentrations of cetuximab for 48 h, and the cell viability was detected using an MTT assay. (e) The SNU-61 cells were transfected with CON and MEIS3 cDNA, and the expression of MEIS3 at the protein level was detected by western blot after 48 h ( n = 3). (f) The SNU-61 CON and MEIS3 groups were treated with different concentrations of cetuximab for 48 h, and PI staining was used to detect changes in the cell cycle. (g) The SNU-61 CON and MEIS3 groups were treated with different concentrations of cetuximab for 48 h, and cell viability was detected using an MTT assay. (h) The LOVO and LOVO MEIS3 shRNA cells were treated with 0.5 mu g/mL cetuximab for 48 h while the SNU-61 and SNU-61 MEIS3 were treated with 0.125 mu g/mL cetuximab for 48 h, and the change in apoptosis was detected using the Annexin V/PI double staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Colon cancer cell lines (a) LOVO-CON, (b) LOVO MEIS3 shRNA, (c) SNU-61 CON, and (d) SNU-61 MEIS3 were treated with different concentrations of cetuximab for 48 h. The expression of EGFR and p-EGFR was detected by western blotting ( n = 3). (e) In LOVO-CON and MEIS3 shRNA cells, expression of MEIS3, p-EGFR, and EGFR was detected by western blot ( n = 3). (f) In SNU-61 CON and SNU-61 MEIS3 cells, the expression of MEIS3, p-EGFR, and EGFR was detected by western blot ( n = 3).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 (a) The IC50 pattern of LOVO-CON, LOVO MEIS3 shRNA, SNU-61 CON, and SNU-61 MEIS3. (b) The LOVO-CON, LOVO MEIS3 shRNA, SNU-61 CON, and SNU-61 MEIS3 cells were treated with different concentrations of crizotinib, and the proliferation of cells was detected using an MTT assay. (c) The LOVO cells were treated with different concentrations of crizotinib, and the expression of MEIS3, p-c-Met, and c-Met was determined by western blot ( n = 3). (d) The LOVO-CON, LOVO MEIS3 shRNA, SNU-61 CON, and SNU-61 MEIS3 cells were treated with different concentrations of miltefosine, and cell proliferation was determined using an MTT assay. (e) The LOVO cells were treated with different concentrations of miltefosine, and the expression of MEIS3, p-Akt, and Akt at the protein level was determined by western blot ( n = 3). (f) LOVO-CON and LOVO MEIS3 shRNA cells were treated with different concentrations of crizotinib and miltefosine, and expression of MEIS3, p-Akt, and Akt at the protein level was determined by western blot ( n = 3).

PA5-61288