rps15a  (Boster Bio)

Journal: FEBS Open Bio

Article Title: Characterization of ribosome heterogeneity during endothelial to hematopoietic transition

doi: 10.1002/2211-5463.70078

Figure Lengend Snippet: The expression pattern of ribosomal proteins during EHT at the proteome level. (A) The distribution and quantity of RPL, RPS and RAP proteins were analyzed using bulk‐cell proteomic data during EHT (EC: endothelial cell, n = 3; HEC: hemogenic endothelial cell, n = 3; HC: hematopoietic cell, n = 3). (B) The average expression level of RPL, RPS and RAP proteins during EHT. (C) The stage‐specific ribosome‐related proteins during EHT. The relative fold change of average protein expression level was represented between adjacent populations. (D, E) The differentially expressed pattern of RPS15a, RPL27 and RACK1 at transcriptome (D) and proteome (E) levels.

Article Snippet: The primary antibodies were as follows: RACK1 (Proteintech, Rosemont, IL, USA; dilution 1 : 200), RPL27 (Proteintech; dilution 1 : 50), RPS15a (Boster, Pleasanton, CA, USA; dilution 1 : 50) and RPS6 (Proteintech; dilution 1 : 50).

Techniques: Expressing

Journal: FEBS Open Bio

Article Title: Characterization of ribosome heterogeneity during endothelial to hematopoietic transition

doi: 10.1002/2211-5463.70078

Figure Lengend Snippet: Quantitative analysis of ribosomal protein expression by immunofluorescence. (A) Representative immunofluorescence images showing subcellular localization of RACK1, RPL27, RPS15a and RPS6 in AECs (CD41 − CD43 − CD45 − CD31 + CD201 − Kit − CD44 + ), HECs (CD41 − CD43 − CD45 − CD31 + CD201 + Kit + CD44 + ) and HCs (CD45 + Kit + ). Scale bars = 10 μm. (B) Quantitative analysis of ribosomal protein expression through immunofluorescence signal intensity measurement. Data were collected from single AEC ( n = 6), HEC ( n = 6) and HC ( n = 5). Data are presented as the mean ± SD and analyzed by unpaired two‐tailed Student's t ‐test. Data were collected from three independent experiments.

Article Snippet: The primary antibodies were as follows: RACK1 (Proteintech, Rosemont, IL, USA; dilution 1 : 200), RPL27 (Proteintech; dilution 1 : 50), RPS15a (Boster, Pleasanton, CA, USA; dilution 1 : 50) and RPS6 (Proteintech; dilution 1 : 50).

Techniques: Expressing, Immunofluorescence, Two Tailed Test

Journal: Clinical and Translational Medicine

Article Title: Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer

doi: 10.1002/ctm2.70249

Figure Lengend Snippet: RPS15AP12‐lncRNA competitively combines miR‐96‐3p to positively regulate RPS15A expression. (A) Spearman's correlation analysis between RNA levels of RPS15AP12 and RPS15A according to the TCGA ovarian cancer cohort. (B) RT‐qPCR assays detecting the expression of RPS15A upon RPS15AP12 KO in OVCAR3 and SKOV3 cell lines. (C) Western blot detecting the protein level of RPS15A upon RPS15AP12 KO in OVCAR3 and SKOV3 cell lines. (D) Representative IHC staining images and quantitative analysis of RPS15A in xenograft tumours from RPS15AP12 knockout and control cells treated nude mice. Scale bar, 100 µm. Statistical analyses showed the IHC staining of RPS15A from xenograft tumours. (E) AGO2 RIP‐PCR detecting the binding of RPS15A and RPS15AP12 with miRNAs. (F) Venn plot showing co‐binding miRNAs shared by RPA15A and RPS15AP12. (G) Schematic diagram of the binding site and sequence in miR‐96, RPS15AP12 and RPS15A. RT‐qPCR detecting miR‐96‐3p level upon miR‐96‐3p mimics and miR‐96‐3p inhibitor in OVCAR3 and SKOV3 cell lines. (H) RT‐qPCR detecting miR‐96‐3p expression upon miR‐96‐3p mimics and miR‐96‐3p inhibitor in OVCAR3 and SKOV3 cell lines. (I) RT‐qPCR detecting RPS15A mRNA level upon miR‐96‐3p mimics and miR‐96‐3p inhibitor in OVCAR3 and SKOV3 cell lines. (J) RT‐qPCR detecting RPS15A mRNA level was performed with control, RPS15AP12‐KO, and RPS15AP12‐KO+miR‐96‐3p inhibitor in OVCAR3 and SKOV3 cell lines. RT‐qPCR detecting RPS15A mRNA level was performed with control, RPS15AP12‐OE, and RPS15AP12‐OE+miR‐96‐3p mimics in OVCAR3 and SKOV3 cell lines. (K) The schematic diagram of wild‐type and mutant 3′UTR of RPS15A for luciferase assays. (L, M) Luciferase assays of RPS15A‐WT and RPS15A‐MUT upon control, miR‐96‐3p mimics and miR‐96‐3p mimics+RPS15AP12‐OE in HEK293T and OVCAR3 cells. One‐way ANOVA, * p < .05, ** p < .01, *** p < .001; NS, not significant.

Article Snippet: Primary antibodies were as follows: METTL3 (1:1000, 15073‐1‐AP, Proteintech), FTO (1:1000, 27226‐1‐AP, Proteintech), YTHDF2 (1:2000, 24744‐1‐AP, Proteintech), RPS15A (1:1000, TA369533S, ORIGENE), MDA5 (1:2000, 21775‐1‐AP, Proteintech), RIG‐1 (1:1000, 20566‐1‐AP, Proteintech), TBK1 (1:1000, #3504, CST), p‐TBK1 (1:1000, 82383‐1‐RR, Proteintech), IRF3 (1:5000, 11312‐1‐AP, Proteintech), p‐IRF3 (1:1000, 29528‐1‐AP, Proteintech), GAPDH (1:10000, 60004‐1‐Ig, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Knock-Out, Control, Binding Assay, Sequencing, Mutagenesis, Luciferase

Journal: Clinical and Translational Medicine

Article Title: Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer

doi: 10.1002/ctm2.70249

Figure Lengend Snippet: RPS15AP12 inhibits the anti‐tumour effect of innate immune‐related pathways in ovarian cancer. (A) Volcano plot showing log2(fold change) and p ‐value of RNA‐seq upon RPS15A knockdown and RPS15AP12 knockout. (B) Overlapped DEGs of RNA‐seq upon RPS15A knockdown and RPS15AP12 knockout. (C) Enrichment network showing innate immune‐related pathways and corresponding hub genes by RPS15A knockdown and RPS15AP12 knockout. (D) Heatmap of 25 shared DEGs in innate immune‐related and proliferation‐related pathways in RNA‐seq upon RPS15A knockdown and RPS15AP12 knockout. (E) RT‐qPCR detecting RNA level of 25 shared DEGs in innate immune‐related and apoptosis‐related pathways in OVCAR3 and SKOV3 cell lines upon RPS15AP12 KO. (F) RT‐qPCR detecting RNA level of 25 shared DEGs in innate immune‐related pathways in OVCAR3 and SKOV3 cell lines upon RPS15A knockdown. (G) Western blot assays detecting protein expression using antibodies as indicated in OVCAR3 and SKOV3 cell lines upon RPS15A knockdown. (H) Western blot assays detecting protein expression using antibodies as indicated in OVCAR3 and SKOV3 cell lines upon RPS15AP12 depletion. One‐way ANOVA, * p < .05, ** p < .01, *** p < .001; NS, not significant.

Article Snippet: Primary antibodies were as follows: METTL3 (1:1000, 15073‐1‐AP, Proteintech), FTO (1:1000, 27226‐1‐AP, Proteintech), YTHDF2 (1:2000, 24744‐1‐AP, Proteintech), RPS15A (1:1000, TA369533S, ORIGENE), MDA5 (1:2000, 21775‐1‐AP, Proteintech), RIG‐1 (1:1000, 20566‐1‐AP, Proteintech), TBK1 (1:1000, #3504, CST), p‐TBK1 (1:1000, 82383‐1‐RR, Proteintech), IRF3 (1:5000, 11312‐1‐AP, Proteintech), p‐IRF3 (1:1000, 29528‐1‐AP, Proteintech), GAPDH (1:10000, 60004‐1‐Ig, Proteintech).

Techniques: RNA Sequencing, Knockdown, Knock-Out, Quantitative RT-PCR, Western Blot, Expressing