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(a) Quantification of the <t>CD44v6-APC-positive</t> population in CPP1 (n=7) and CTC44 (n=5) cells transfected with miR-4745-3p ( miR4745 ) or miRVana microRNA Mimic Negative Control #1 (miCTRL) mimics (right panel). Percentage of CD44v6-APC positive CPP1 cells is indicated in inset boxes for a representative experiment (left panel). (b) Representative images of tumorspheres (upper panel) and the percentage (lower panel) of sphere-forming cells in patient-derived CPP1 and circulating tumor (CTC44) colon cancer cells transfected with miR-4745-3p ( miR4745 ) or miCTRL mimics. Data from individual replicates (n=15 per experiment) are presented, along with the mean ± SEM from six independent experiments. (c) Percentage of sphere-forming cells in patient-derived CPP1 colon cancer cells transfected with miR4745 or negative control (miCTRL) mimics and, maintained as first-generation colonospheres (S1) or after several passages (S2, S3). Data from individual replicates (n=12 per experiment) are presented, along with the mean ± SEM from three independent experiments. (d) Percentage of surviving cancer stem cells (Aldefluor-positive) measured 48 hours following treatment with specified concentrations of FIRI (1X = 50 µM 5-FU + 500 nM SN38) in vitro . Sorted CPP1 Aldefluor-positive cells were first transfected with 50 nM miR-4745-3p ( miR-4745 ) or miRVana microRNA Mimic Negative Control #1 (miCTRL) mimics, 24 hours prior to FIRI treatment. Data are expressed as mean ± SEM (n = 3). (e) Percentage of Aldefluor-positive cells (% ALDH + cells) in CPP1 cells transfected with miR-4745 or miCTRL mimics and then exposed for 72 hours to chemotherapy (Firi=5µM 5-FU + 50nM SN38). Data are expressed as mean ± SEM (n=6). *, p<0.05; **, p<0.005, ***, p < 0.001, Mann Whitney test.
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(a) Quantification of the CD44v6-APC-positive population in CPP1 (n=7) and CTC44 (n=5) cells transfected with miR-4745-3p ( miR4745 ) or miRVana microRNA Mimic Negative Control #1 (miCTRL) mimics (right panel). Percentage of CD44v6-APC positive CPP1 cells is indicated in inset boxes for a representative experiment (left panel). (b) Representative images of tumorspheres (upper panel) and the percentage (lower panel) of sphere-forming cells in patient-derived CPP1 and circulating tumor (CTC44) colon cancer cells transfected with miR-4745-3p ( miR4745 ) or miCTRL mimics. Data from individual replicates (n=15 per experiment) are presented, along with the mean ± SEM from six independent experiments. (c) Percentage of sphere-forming cells in patient-derived CPP1 colon cancer cells transfected with miR4745 or negative control (miCTRL) mimics and, maintained as first-generation colonospheres (S1) or after several passages (S2, S3). Data from individual replicates (n=12 per experiment) are presented, along with the mean ± SEM from three independent experiments. (d) Percentage of surviving cancer stem cells (Aldefluor-positive) measured 48 hours following treatment with specified concentrations of FIRI (1X = 50 µM 5-FU + 500 nM SN38) in vitro . Sorted CPP1 Aldefluor-positive cells were first transfected with 50 nM miR-4745-3p ( miR-4745 ) or miRVana microRNA Mimic Negative Control #1 (miCTRL) mimics, 24 hours prior to FIRI treatment. Data are expressed as mean ± SEM (n = 3). (e) Percentage of Aldefluor-positive cells (% ALDH + cells) in CPP1 cells transfected with miR-4745 or miCTRL mimics and then exposed for 72 hours to chemotherapy (Firi=5µM 5-FU + 50nM SN38). Data are expressed as mean ± SEM (n=6). *, p<0.05; **, p<0.005, ***, p < 0.001, Mann Whitney test.

Journal: bioRxiv

Article Title: A Novel miR-4745 -KLC2 Axis Regulates Cancer Stem Cell Traits in Colorectal Cancer

doi: 10.64898/2026.01.07.697660

Figure Lengend Snippet: (a) Quantification of the CD44v6-APC-positive population in CPP1 (n=7) and CTC44 (n=5) cells transfected with miR-4745-3p ( miR4745 ) or miRVana microRNA Mimic Negative Control #1 (miCTRL) mimics (right panel). Percentage of CD44v6-APC positive CPP1 cells is indicated in inset boxes for a representative experiment (left panel). (b) Representative images of tumorspheres (upper panel) and the percentage (lower panel) of sphere-forming cells in patient-derived CPP1 and circulating tumor (CTC44) colon cancer cells transfected with miR-4745-3p ( miR4745 ) or miCTRL mimics. Data from individual replicates (n=15 per experiment) are presented, along with the mean ± SEM from six independent experiments. (c) Percentage of sphere-forming cells in patient-derived CPP1 colon cancer cells transfected with miR4745 or negative control (miCTRL) mimics and, maintained as first-generation colonospheres (S1) or after several passages (S2, S3). Data from individual replicates (n=12 per experiment) are presented, along with the mean ± SEM from three independent experiments. (d) Percentage of surviving cancer stem cells (Aldefluor-positive) measured 48 hours following treatment with specified concentrations of FIRI (1X = 50 µM 5-FU + 500 nM SN38) in vitro . Sorted CPP1 Aldefluor-positive cells were first transfected with 50 nM miR-4745-3p ( miR-4745 ) or miRVana microRNA Mimic Negative Control #1 (miCTRL) mimics, 24 hours prior to FIRI treatment. Data are expressed as mean ± SEM (n = 3). (e) Percentage of Aldefluor-positive cells (% ALDH + cells) in CPP1 cells transfected with miR-4745 or miCTRL mimics and then exposed for 72 hours to chemotherapy (Firi=5µM 5-FU + 50nM SN38). Data are expressed as mean ± SEM (n=6). *, p<0.05; **, p<0.005, ***, p < 0.001, Mann Whitney test.

Article Snippet: CD44v6-APC (clone REA706, 130-111-238; Macs Miltenyi) antibody was incubated with 100 000 cells in PBS containing 5% FBS for 20 min at 4°C.

Techniques: Transfection, Negative Control, Derivative Assay, In Vitro, MANN-WHITNEY