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Image Search Results
Journal: Stem Cell Reports
Article Title: CCR5 Signaling Promotes Murine and Human Hematopoietic Regeneration following Ionizing Radiation
doi: 10.1016/j.stemcr.2019.04.023
Figure Lengend Snippet: CCL5 Promotes Cell Cycling and Cell Survival after Irradiation (A) Left, representative flow cytometric analysis of KSL cells in G0 (Ki67 − 7-AAD − ), G 1 (Ki67 + 7-AAD − ), and G 2 /S/M (Ki67 + 7-AAD + ) phase at 72 h following 300 cGy in culture with CCL5 + TSF (CCL5) compared with TSF alone. (B) Cell-cycle analysis of KSL cells at 72 h following 300 cGy in culture with CCL5 + TSF (CCL5) compared with TSF alone. n = 7–8 per group, ∗ p = 0.009 and p = 0.005 for G0/G1 and G2/S/M, respectively. (C) RT-PCR analysis of Cdk2 , Cdk4 , and Cdk6 mRNA expression from KSL cells at 72 h following 300 cGy and treatment with TSF or 30 ng/mL CCL5. n = 3 per group, ∗ p = 0.0002, p = 0.004, and p = 0.004 for Cdk2 , Cdk4 , and Cdk6 , respectively. Data are normalized to GAPDH and TSF for each Cdk . (D) Schematic diagram of study design for E–6G. At 24 h after 500 cGy TBI, mice were injected with saline or 0.1 μg/g body weight of CCL5 and analyzed at 2 h postinjection. (E) RT-PCR analysis of Cdk2 , Cdk4 , and Cdk6 of BM Lin − cells. n = 4 per group, ∗ p = 0.003, p < 0.0001, and p < 0.0001 for Cdk2 , Cdk4 , and Cdk6 , respectively. Data are normalized to GAPDH and saline for each Cdk . (F) Representative flow cytometric Ki67/7-AAD cell-cycle analysis from KSL cells following 500 cGy TBI and treatment with CCL5 or saline. Nonirradiated control is shown for comparison. (G) Levels of G0/G1 and G2/S/M of KSL cells from irradiated CCL5- or saline-treated C57BL/6 mice. n = 5 per group, ∗ p < 0.001 for G0/G1 and G2/S/M. (H) Left, representative flow cytometric analysis of annexin V + KSL cells and progeny at 72 h following 300 cGy in culture with CCL5 (blue line) compared with TSF alone (gray line). Right, quantification of total annexin V + cells day 7 following 300 cGy and treatment with CCL5 or TSF alone. n = 3 per group, ∗ p < 0.0001. (I) Representative flow cytometric analysis and quantification of γ-H2AX in KSL and progeny following 300 cGy irradiation and 24 h culture with CCL5 (blue) or TSF alone (gray). Isotype is shown as black curve. n = 6 per group, ∗ p = 0.02. (J) Flow cytometric analysis of annexin V + from KSL cells and progeny of Ccr5 +/+ and Ccr5 −/− mice at 72 h following 300 cGy. n = 6 per group, ∗ p = 0.02. Data are shown as means ± SEM. Student's t test (two-tailed with unequal variance) was applied to the data.
Article Snippet: RT-PCR for Cdk2 , Cdk4 , and Cdk6 was performed on irradiated KSL cells or BM Lin − cells as specified in the figure legends (
Techniques: Irradiation, Cell Cycle Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Injection, Saline, Control, Comparison, Two Tailed Test