Journal: eLife

Article Title: Transcriptomic analyses reveal rhythmic and CLOCK-driven pathways in human skeletal muscle

doi: 10.7554/eLife.34114

Figure Lengend Snippet:

Article Snippet: Chemical compound, drug , 17:0-14:1 PE , Avanti Polar Lipids , LM1104 , .

Techniques: Western Blot, Recombinant, Sequencing, Picogreen Assay, Activity Assay, Transfection, Software

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Closed-system manufacturing of therapeutic NK cells using automated cell enrichment and concentration processes enables scalable, robust and cost-effective solutions

doi: 10.3389/fbioe.2025.1586912

Figure Lengend Snippet: NK drug substance composition before and after the concentration process using the CliniMACS Prodigy ® . (A) Total CD45 + cell (identified as CD45 + living) number before and after the cell concentration process (n = 29). (B) Total CD45 + cell yield after the cell concentration process, according to low (n = 7), medium (n = 14) or high (n = 8) cell culture volume. (C) CD56 + NK cell (identified as CD45 + CD56 + CD3 − living) purity before and after cell concentration process (n = 29). (D) CD56 + cell purity after cell concentration process, according to low (n = 7), medium (n = 14) or high (n = 8) cell culture volume. (E) CD19 + B cell impurity content (identified as CD45 + CD19 + living) before and after cell concentration process (n = 7). For all plots, the central line denotes the median; boxes delimitate the 25th and 75th percentiles and whiskers mark the minimum and maximum. For statistical analysis, paired T-test was used in (A, C, E) , one-way ANOVA in (B,D) .

Article Snippet: CD19 , PE , LT19 , 170-081-057 , Miltenyi , GMP 1-22, R&D 25-36.

Techniques: Concentration Assay, Cell Culture

Journal: RNA nanomed

Article Title: CD133-Guided RNA Nanoparticle Delivery of FTO siRNA Impairs Leukemia Resistance to Tyrosine Kinase Inhibitor Therapy

doi: 10.59566/isrnn.2025.0201e

Figure Lengend Snippet: Expression and regulation of LSC markers in naïve, larger clones and TKI resistant leukemia cells. (A) Quantification of CD25+ cells in K562 cells treated with 0.3 or 1 μM of nilotinib for 48 hours, as measured by flow cytometry. (B) Left: Graphs representing colony numbers of parental and nilotinib-resistant K562 cells cultured in drug-free medium for 72 hours; Right: Flow cytometry analysis depicting the percentage of CD25+ cells in large vs small clones. (C) Graphs showing quantification of CD25+, CD44+ and CD133+ cells measured in parental vs NIR or IMR K562 cells, as measured by flow cytometry. (D) qPCR measuring indicated LSC marker expression in parental vs resistant Kasumi-1, MV4–11 and K562 cells. (E) Western blot and quantification of KIT (CD117) and EZH2 protein expression in parental, NIR-resistant, and IMR-resistant K562 cells. Band intensity was quantified and normalized to a loading control (β-actin). (F-G) qPCR measuring changes in the indicated stem cell markers in FTO knockdown (F) or FTO inhibitor CS2-treated (G) K562 cells resistant to nilotinib. (H) m 6 A immunoprecipitation (IP) was performed in mRNA from K562 resistant cells. The eluted RNA was converted to cDNA and transcript levels of KIT, CD25 and CD44 were measured by qPCR. Data are representative of three independent experiments. Par, parental; NIR, nilotinib resistance; IMR, imatinib resistance; Kas, Kasumi-1; shctrl, scramble control; * P <0.05, ** P <0.01, *** P <0.001; ns, not statistically significant.

Article Snippet: To measure the percentage of LSCs, different TKI resistant and parental cells were stained with antibodies against CD44 (Miltenyi Biotec, 130-113-334), CD25 (Miltenyi Biotec, 170-081-060) or CD133 (Miltenyi Biotec, 170-081-001).

Techniques: Expressing, Clone Assay, Flow Cytometry, Cell Culture, Marker, Western Blot, Control, Knockdown, Immunoprecipitation

Journal: RNA nanomed

Article Title: CD133-Guided RNA Nanoparticle Delivery of FTO siRNA Impairs Leukemia Resistance to Tyrosine Kinase Inhibitor Therapy

doi: 10.59566/isrnn.2025.0201e

Figure Lengend Snippet: Strong uptake of CD133-targeted RNA nanoparticle and efficient knockdown of FTO in TKI resistant CML cells. (A) Design of 3WJ-CD133apt nanoparticles with Alexa-647 dye. (B-D) K562 resistant cells were stained by 50 nM of the indicated nanoparticles in FBS-free RPMI 1640 medium for 1 hour, and the labeled cells were detected by flow cytometry (B) and visualized by confocal microscope (C). Graphs are quantification of RNA nanoparticle-bound cells (D). Data represents three independent experiments. (E-F) Cartoon images showing the designs of RNA nanoparticles. (G) CD133-targeted RNA nanoparticles carrying FTO siRNA impair the expression of FTO gene in resistant CML cells. Imatinib resistant K562 cells were treated with 100 μM FTO siRNA-loaded RNA nanoparticles for 48 or 72 hours, and FTO expression was detected by qPCR. Data represents biological and technical duplicates. Note: 1, 3WJ; 2, 3WJ-CD133-siFTOScr-AF647; 3, 3WJ-CD133-siFTO-AF647; A15, 3WJ-CD133A15-AF647; B19, 3WJ-CD133B19-AF647; Scr, scramble; siR, siRNAs. * P <0.05.

Article Snippet: To measure the percentage of LSCs, different TKI resistant and parental cells were stained with antibodies against CD44 (Miltenyi Biotec, 130-113-334), CD25 (Miltenyi Biotec, 170-081-060) or CD133 (Miltenyi Biotec, 170-081-001).

Techniques: Knockdown, Staining, Labeling, Flow Cytometry, Microscopy, Expressing

Journal: RNA nanomed

Article Title: CD133-Guided RNA Nanoparticle Delivery of FTO siRNA Impairs Leukemia Resistance to Tyrosine Kinase Inhibitor Therapy

doi: 10.59566/isrnn.2025.0201e

Figure Lengend Snippet: CD133-guided FTO siRNA nanoparticles inhibit growth in nilotinib-resistant K562 cells. Nilotinib-resistant K562 cells were treated with either 3WJ-CD133-siFTO-AF647 (si-FTO) or a control 3WJ-CD133-siFTOScr-AF647 (SCR). (A) qPCR analysis shows the expression of indicated m6A regulators at the RNA level. Data represents three independent biological replicates. (B) Western blot analysis shows the protein expression of the indicated m 6 A regulators. The blot shown is representative of triplicate drug treatment experiments. (C-D) Colony-forming analysis demonstrated the impaired proliferation of resistant cells post-treatment with si-FTO compared to SCR. C, the shown images are the overview of colony formation (4 ×; triplicate wells) and the representative colony (20 ×; right); D, graph is the quantification of colony number in three wells. The data shown is the average of triplicates with ±SEM. *** P <0.001. (E) Spheroid-forming assays reveal the inhibition of resistant cell growth by si-FTO compared to SCR. The shown images are the overview of spheroid formation (4 ×; triplicate wells) and the representative spheroids (40 ×; right).

Article Snippet: To measure the percentage of LSCs, different TKI resistant and parental cells were stained with antibodies against CD44 (Miltenyi Biotec, 130-113-334), CD25 (Miltenyi Biotec, 170-081-060) or CD133 (Miltenyi Biotec, 170-081-001).

Techniques: Control, Expressing, Western Blot, Inhibition