EMBO/soda-nel huggingface dataset:
Multi-modal meta-analysis of cancer cell line omics profiles identifies ECHDC1 as a novel breast tumor suppressor (2021)
Figure 5

Figure 5: Identification of ECHDC1 as breast cancer tumor suppressor gene.

(A) The CLIP-signature of ECHDC1 suggests that it was hypermethylated and down-expressed in all the breast cancer cell lines (n=24) in which it was identified as rCCS gene.

(B) Breast cancer specific survival (BCSS) based on gene expression and methylation levels of ECHDC1 in breast cancer patient tumors in the combined Metabric and Oslo datasets (n=3885). Patients in the low GEXP category class have lower BCSS than those in the non-low GEXP group. Numbers above the x-axis line indicate the number of patients in each group, defined by the color code, at each time point. P-value from age-adjusted Cox-proportion hazard model.

(C) Benign breast epithelial MCF10A and breast carcinoma BT-474 cells were embedded in 3D collagen as single cells or as spheroids, respectively, and the growth was followed for 5 days. Light micrographs show filamentous actin (phalloidin) and nuclei (Hoechst) in representative cell colonies. Quantitative assessment of the nuclei counts per colony show the induced proliferation in MCF10A cells after ECHCD1 sgRNA knockout. At 72 hours, MCF10A mock versus ECHDC1_sgRNA_1 and ECHDC1_sgRNA_2 P < 0.05; at 96 hours mock versus ECHDC1_sgRNA_1, ECHDC1_sgRNA_2 and ECHDC1_sgRNA_3 P < 0.001; at 120 hours mock versus ECHDC1_sgRNA_1, ECHDC1_sgRNA_2 and ECHDC1_sgRNA_3 P < 0.0001. Nuclei count relative to mock 0 hours. Error bars indicate mean ± SEM; n ≥ 10 colonies. Statistical significance was assessed with one-way ANOVA with Tukey's multiple comparison test. Scale bar 50 µm.

(D) Metabolic pathway of propanoate metabolism.

(E) Measured metabolite levels of intermediates in propanoate metabolism in select breast cancer cell lines with or without the ECHDC1 rCCS status (n=7 in both groups). Boxes represent the interquartile range, whiskers represent the range of the values and solid line within the box correspond to the median value. Outlier points indicates values not included between the whiskers. Statistical significance was assessed with Wilcoxon test.
Multi-modal meta-analysis of cancer cell line omics profiles identifies ECHDC1 as a novel breast tumor suppressor (2021)
Figure 6

Figure 6: Identification of novel synthetic lethal interactions.

(A) Proportion of KRAS mutated (Mut) and KRAS wild type (WT) cancer cell lines with KRAS identified as a rCCS gene. P-value was calculated with Fisher's exact test.

(B) The modalities that support the rCCS status of KRAS and the proportion of cell lines having that evidence in the KRAS mutated cell lines (n=155).

(C) Proportion of PIK3CA mutated (Mut) and PIK3CA wild type (WT) cancer cell lines with PIK3CA identified as a rCCS gene. P-value was calculated with Fisher's exact test.

(D) The modalities that support the rCCS status of PIK3CA and the proportion of cell lines having that evidence in the PIK3CA mutated cell lines (n=140).

(E) Systematic identification of cancer driver genes specific to epithelial cancer cell lines (n=737) under multiple settings of CLIP run. rCCS genes identified by CLIP are enriched for known cancer drivers compared to non-driver genes, even after excluding the FUNC data modality from the CLIP approach. Boxes represent the interquartile range, whiskers represent the range of the values and solid line within the box correspond to the median value. Statistical significance was assessed with Wilcoxon test.

(F) Proportion of PTEN mutated (Mut) and PTEN wild type (WT) cancer cell lines in which DDX27 was identified as a rCCS gene. P-value was calculated with Fisher's exact test.

(G) The modalities that supported the rCCS status of DDX27 and the proportion of cell lines having that evidence in the PTEN mutated cell lines (n=77).

(H) Survival analysis based on mRNA expression levels of DDX27 in patients with endometrial cancer in the TCGA dataset. Expression levels were divided into 2 classes, high (n=203) and low (n=322), based on mean expression level of DDX27 (logFPKM=18.27). Patients in the high class showed lower survival probability than those in the low class (P=.4.2×10-4; log-rank test).

(I) mRNA expression levels of DDX27 in PTEN mutated (Mut, n=302) and PTEN wild type (WT, n=224) endometrial patient tumors in the TCGA dataset. Triangles correspond to the median value. P-value was calculated with Wilcoxon test.
TEFM regulates both transcription elongation and RNA processing in mitochondria (2019)
Figure 1

Figure 1 Disruption of Tefm in the germline and heart. (A) Targeting strategy for disruption of the Tefm gene. LoxP sites flanking exon 2 of the Tefm gene together with puromycin selection marker (PuroR) were inserted into the mouse genome by homologous recombination. The PuroR cassette was excised by mating with Flp-recombinase mice to obtain heterozygous floxed Tefm mice (Tefm+/loxP). Whole-body and tissue-specific Tefm knockout mice were obtained by crossing Tefm+/loxP with different Cre-recombinase mice. (B) Morphology of wild-type (Tefm+/+) and Tefm homozygous knockout (Tefm-/-) embryos at embryonic day 8.5 (analyzed embryos, n= 43). Scale bar, 500 μm. (C) Western blot analyses of the TEFM protein level in heart mitochondrial extracts from 8-week-old control (L/L) and tissue-specific Tefm knockout (Tefm knockout; L/L, cre) mice (n = 12 mice for each group). VDAC was used as loading control. (D) Survival curve for control (L/L; n= 60) and Tefm knockout (L/L, cre; n= 22) mice. (E) Heart weight to body weight ratio in 4- and 8-week-old control (L/L) and Tefm knockout mice (L/L, cre). At 4 weeks: L/L n= 22, L/L, cre n= 15; 8 weeks: L/L n= 41, L/L, cre n= 39.

Data information: In (E), data are presented as mean ± SEM. ***P<0.001; Student's t-test. 4 weeks P= 5.041476e-009, 8 weeks P= 1.096535e-033.
TEFM regulates both transcription elongation and RNA processing in mitochondria (2019)
Figure 2

Figure 2 Knockout of Tefm in heart causes mitochondrial dysfunction. (A) Transmission electron micrographs of heart tissue in 8-week-old control and Tefm knockout mice (n = 5 mice for each group). Scale bar, 2 μm (upper panel) and 1 μm (lower panel). (B) Southern blot analyses of mtDNA levels in control and Tefm knockout hearts at the age of 4 and 8 weeks (Upper 2 blots; n= 6 mice at each time-point for each group). 18S rDNA was used as loading control. DNA isolated from Lrpprc and Mterf4 knockout hearts used as controls. Western blot analyses of VDAC and Histone H3 protein levels in control and Tefm knockout hearts at the age of 4 and 8 weeks (Bottom 2 blots; ; n= 6 mice at each time-point for each group). VDAC represents mitochondria loading and Histone H3 was used as a loading control for nucleus DNA. (C) Respiratory chain complex activities were measured spectrophotometrically and normalized to citrate synthase activity in heart mitochondria from 8-week-old control and Tefm knockout mice (n= 5 mice for each group). The analyzed enzyme activities are NADH coenzyme Q reductase (complex I), NADH cytochrome c reductase (complex I/III), succinate dehydrogenase (complex II) and cytochrome c oxidase (complex IV). (D) Barplot of the significantly changed proteins (Benjamini-Hochberg adjusted P <0.05) in Tefm knockout mice compared to control mice (n= 5 mice for each group), organized by individual OXPHOS complexes. Proteins are organized in alphabetic order based on gene name. Bars represent mean log2 [L/L, cre / L/L] ± 95% confidence interval. (E) Boxplots of the protein intensity (LFQ) of TEFM (top) and POLRMT (bottom). Red dot: mean; blue dots: intensity value for each biological replicate (n= 5 mice for each group). (F) Western blot analyses of POLRMT protein levels in control and Tefm knockout heart mitochondria at the age of 8 weeks (n= 12 mice for each group). VDAC was used as loading control.

Data information: In (C), data are presented as mean ± SEM. ***P<0.001; Student's t-test.

CI P= 4.27443E-06, CI/CIII P= 2.0553E-05, CII P= 2.76012E-05, CII/CIII P= 2.00732E-07, CIV P= 1.72156E-06.
TEFM regulates both transcription elongation and RNA processing in mitochondria (2019)
Figure 3

Figure 3 Loss of TEFM impairs mtDNA transcription elongation and decreases steady-state levels of promotor-distal transcripts. (A) Schematic linear map of mouse mtDNA indicating the relative position of mtRNA transcripts analyzed by northern blot. (B-E) Northern blot analyses (B and D) and relative quantification (C and E) of mtRNA transcripts in control and Tefm knockout mouse hearts at the age of 4 and 8 weeks (n= 12 mice at each time-point for each group). 18S rRNA and 5.8S rRNA were used as loading controls, respectively.

Data information: In (C and E), data are presented as mean ± SEM. *P<0.05, **P<0.01 and ***P<0.001; Student's t-test. (C) HSP transcripts: 12S rRNA 4 weeks P= 0.005966044, 8 weeks P= 0.0806819; 16S rRNA 4 weeks P= 0.000223004, 8 weeks P= 0.00243443; Nd1 4 weeks P= 0.0352088, 8 weeks P= 0.000344158; Cox1 4 weeks P= 0.000353895, 8 weeks P= 0.00145773; Cox2 4 weeks P= 0.0145193, 8 weeks P= 0.00578484; Cox3 4 weeks P= 0.00834317, 8 weeks P= 0.000392364; Nd5/Cyb 4 weeks P= 0.0010185, 8 weeks P= 0.00153049; Nd5 4 weeks P= 0.024621, 8 weeks P= 7.38074E-05; Cyb 4 weeks P= 3.46175E-06, 8 weeks P= 0.000991942; LSP transcript: Nd6 4 weeks P= 0.000162704, 8 weeks P= 0.000187855. (E) HSP transcripts: Tf 4 weeks P= 0.0297685, 8 weeks P= 0. 000212544; Tv 4 weeks P= 0.842023, 8 weeks P= 0.269231; Tl1 4 weeks P= 0.00921072, 8 weeks P= 0.000430068; Tm 4 weeks P= 0.0084742, 8 weeks P= 0.000142524; Tk 4 weeks P= 0.00297015, 8 weeks P= 0.00010425; Ts2 4 weeks P= 0.0128829, 8 weeks P= 2.27321E-05; Tl2 4 weeks P= 0.00701324, 8 weeks P= 0.000411549; Tt 4 weeks P= 0.00572118, 8 weeks P= 0.000807943; LSP transcript: Tp 4 weeks P= 0.0427191, 8 weeks P= 0.0483328; Te 4 weeks P= 0.464031, 8 weeks P= 0.13412; Ts1 4 weeks P= 0.0382215, 8 weeks P= 0.000478237; Tc 4 weeks P= 0.00418795, 8 weeks P= 0.000287525; Tn 4 weeks P= 0.00531668, 8 weeks P= 0.000374367; Ta 4 weeks P= 0.00247257, 8 weeks P= 0.000564597; Tq 4 weeks P= 0.00385524, 8 weeks P= 0.000727981.
TEFM regulates both transcription elongation and RNA processing in mitochondria (2019)
Figure 4

Figure 4 Accumulated shorter LSP transcripts in TEFM-depleted mitochondria do not contribute to mtDNA replication. (A) In organello transcription analyses using heart mitochondria isolated from 8-week-old control and Tefm knockout mice (n= 5 mice for each group). The steady-state levels of 12S rRNA and tRNAV were used as endogenous controls, due to their unchanged levels. VDAC was used as loading control for the equal input of mitochondrial extracts. This experiment has performed five times with either two or three replicates. (B) Northern blot analyses of 7S RNA level in 4- and 8-week-old control and Tefm knockout mouse hearts (n= 12 mice at each time-point for each group). 18S rRNA was used as loading control. RNA from tissue-specific Mterf4 knockout mouse hearts and tissue-specific Polrmt knockout mouse hearts were loaded as positive and negative control, respectively. (C) Southern blot analyses of mtDNA to obtain 7S DNA in 8-week-old control and Tefm knockout mice. Heart mitochondria from 3 control or knockout mice were pooled for preparation of mtDNA. Equal amounts of mtDNA shown on the top of the gel, were loaded as the endogenous control. This experiment has performed three times. (D) In organello replication assays were performed on heart mitochondria isolated from 8-week-old control and Tefm knockout mice (n= 5 mice for each group). VDAC was used as loading control. (E) Northern blot analyses of the 7S RNA levels in 4- and 8-week-old control and Tefm knockout mouse hearts (n= 12 mice for each group). 5S rRNA was used as loading control. RNA from tissue-specific Mterf4 knockout mouse hearts and tissue-specific Polrmt knockout mouse hearts were loaded as positive and negative control, respectively. (F) Mapping 3' termini of mitochondrial 7S RNA/early LSP transcripts in control (L/L) and Tefm knockout (L/L, cre) mouse hearts at the age of 8 weeks. A 3' RACE was performed using linker-ligated RNA and a forward primer corresponding to the first 45 bp from the transcription initiation site at LSP. CSB, conserved sequence block.
TEFM regulates both transcription elongation and RNA processing in mitochondria (2019)
Figure 5

Figure 5 Transcriptome-wide analysis reveals that mtDNA transcription elongation is linked to RNA processing (A) Transcriptome-wide analysis of the effects of TEFM loss on mtRNA transcripts by TruSeq RNA sequencing. A complete map of mtDNA showing the change in sequence read coverage (log2 fold change [L/L, cre / L/L]) between Tefm knockout (L/L, cre; n= 4) and control (L/L; n= 3) mice on both strands. Increased reads are shown in red and decreased reads in blue. The mtDNA is displayed in the central track, with mt-rRNAs in green, mt-mRNAs in light blue and mt-tRNAs in grey. (B) Strand-specific RT-qPCR detection of the mitochondrial precursor transcripts containing the antisense of mt-tRNAA and antisense of mt-tRNAC in control and Tefm knockout mice (n= 12 mice for each group). The mt-mRNAs are shown in light blue and mt-tRNAs in grey. Arrows show the binding position of RT-qPCR primers. Actin transcript was used as endogenous control. (C) Strand-specific RT-qPCR detection of the mitochondrial precursor transcripts spanning the junction of Nd5 and antisense of Nd6 in control and Tefm knockout mice (n= 25 mice for each group). The mt-mRNAs are shown in light blue. Arrows show the binding position of RT-qPCR primers. Actin transcript was used as endogenous control. (D) Northern blot analyses of the precursor transcripts containing the antisense of mt-tRNAA in 4- and 8-week-old control and Tefm knockout mouse hearts (n= 6 mice for each group).

Data information: In (B and C), data are presented as mean ± SEM. *P<0.05, **P<0.01; Student's t-test. C: P= 0.0089; D: P= 0.0260.
TEFM regulates both transcription elongation and RNA processing in mitochondria (2019)
Figure 6

Figure 6 TEFM interacts with RNA processing factors. Volcano plot showing TEFM-BioID. TEFM-BirA*-HA was stably expressed in HEK 293 cells, followed by streptavidin affinity purification and mass spectrometry analysis to detect the associated proteins. Mitochondrial targeted BirA* (MTS-BirA*) was used as a control. The significantly enriched proteins are presented above the horizontal dashed line with 5% false discovery rate (FDR) significance. The blue dots highlight proteins involved in mtDNA transcription and the green dots represent proteins involved in RNA metabolism. All other proteins are in grey.

Expanded View Figure Legends
Intraflagellar transport proteins 172, 80, 57, 54, 38 and 20 form a stable tubulin-binding IFT-B2 complex (2016)
Figure 1

Figure 1. IFT172, 80, 57, 54, 38 and 20 form an IFT-B2 sub-complex

(A) SDS-PAGE gels stained with coomassie showing the purity of the Chlamydomonas IFT-B2 components, over-expressed in E. coli or insect cells, after the last purification step by SEC. IFT54/20 and IFT57/38 were purified as hetero-dimeric complexes whereas IFT80, IFT172 and an N-terminal IFT172(1-968) (IFT172ΔC) constructs were purified as single components. A cartoon representation of the predicted domain architecture of IFT-B2 components using HHPRED (Söding et al, 2005) is shown at the center.

(B)Pull-down analysis for the hexameric IFT-B2 complex. A mixture of IFT172ΔC, IFT80, and the IFT57/38 complex ('input') shows only weak non-specific binding to GSH-resin ('empty'), but all proteins are pulled down by IFT54/20-GST complex. Note that the IFT20-GST and IFT38 proteins co-migrate on the gel and cannot be properly resolved. However, the presence of the IFT57 protein in the pull-down sample implies that its binding partner IFT38 is also there.

(C) (left) SEC elution profiles of individual IFT-B2 components and of the assembled hexameric IFT-B2 complex. The elution peak of IFT-B2 is shifted compared to all of the individual components. Note that the peak heights were adjusted for better visualization of the result. (right) Coomassie stained SDS PAGE gel of the six main fractions of the IFT-B2 SEC elution peak demonstrating the presence of all six IFT-B2 subunits. Weak contaminating bands are marked with asterisks.
Intraflagellar transport proteins 172, 80, 57, 54, 38 and 20 form a stable tubulin-binding IFT-B2 complex (2016)
Figure 2

Figure 2. Architecture of IFT-B2

(A-C) SEC elution profiles and corresponding SDS PAGE gels of IFT172ΔC, IFT57/38, IFT80, IFT54/20 as well as mixtures of IFT172ΔC/57/38, IFT80/57/38 and IFT172ΔC/80/57/38 demonstrating that IFT57/38 associates with both IFT80 and IFT172ΔC. Only the central peak fraction from each SEC experiment is shown on the SDS PAGE gel. Note that the presence of both IFT172ΔC and IFT80 in a single peak in (c) (peak 3) is due to the fact that these two proteins elute in the same position, and not because they physically interact (see also Appendix Fig. S2B).

(D) GSH resin pull-down of a mixture of IFT80 and IFT172ΔC (input, lane 1) with GST-tagged CH-domains of IFT57 or IFT38 (IFT57CH or IFT38CH) or with the GST-tagged coiled-coil (CC) region of IFT57 mixed with His-tagged coiled-coil region of IFT38. The SDS-PAGE gel shows interaction between IFT57CH-IFT172ΔC, IFT38CH-IFT80 and IFT57CC-IFT38CC.

(E) Ni-NTA resin pull-down of untagged IFT54/20 with His-tagged IFT57/38 (both subunits His-tagged). The two bands for IFT38 on the SDS PAGE gel correspond to His-tagged (upper band) and proteolytically cleaved untagged IFT38 (lower band).

(F) Ni-NTA resin pull-down of untagged IFT54/20 with His-tagged IFT80.

(G) Schematics of the interactions between IFT-B2 components supported by the data in panels a-f of this figure as well as in Appendix Fig. S2. Note that it is currently not known if the coiled-coil interactions between IFT54, 20, 57 and 38 are parallel or anti-parallel.
Intraflagellar transport proteins 172, 80, 57, 54, 38 and 20 form a stable tubulin-binding IFT-B2 complex (2016)
Figure 3

Figure 3. Structure and tubulin-binding of IFT54CH

(A) Ni2+-NTA affinity pull-down of unpolymerised bovine αβ-tubulin with His-tagged IFT-B2 (sub)-complexes. A Coomassie stained SDS PAGE gel shows the IFT-B2 complexes used for pull-downs (top) and a western blot for anti-α-tubulin shows co-precipitated αβ-tubulin (bottom).

(B) Cartoon representation of the crystal structure of CrIFT54CH (left), the electrostatic surface potential (middle), and the amino acid conservation mapped onto the surface of the structure (right).

(C) Cartoon representation of the crystal structure of MmIFT54CH (left) and the electrostatic surface potential (middle).

(D) Superimposition of the CH-domains of CrIFT54CH, MmIFT54CH and CrIFT81 (pdb: 4VLP).

(E) Cartoon representation of the crystal structure of MmIFT54CH domain with the residues of the positively charged patch shown as sticks. Three residues whose mutation to glutamate abolished tubulin-binding are indicated with a red circle.

(F) Ni2+-NTA affinity pull-down of unpolymerised bovine αβ-tubulin with WT and mutant MmIFT54CH. (top) Coomassie stained SDS PAGE gel of input (I) and pulled down (PD) proteins and the corresponding western blot to visualize αβ-tubulin.
Intraflagellar transport proteins 172, 80, 57, 54, 38 and 20 form a stable tubulin-binding IFT-B2 complex (2016)
Figure 5

Figure 5. IFT-B1 and IFT-B2 form a salt-stable complex via an (IFT88/52N)-(IFT57/38) interaction

(A) Coomassie-stained SDS-PAGE gel showing GST-pulldown experiments between the IFT-B2 complex and an IFT88/52N subcomplex. The pre-assembled IFT-B2 complex (lane 1) shows no non-specific interaction with empty GSH-beads (lane 2), but is efficiently pulled down by IFT88/52N-GST (with an IFT52(1-335) construct comprising the N-terminal domain as well as the IFT88-binding region) (lane 3). Both IFT88 and IFT52N are required for the interaction with IFT-B2 because neither the N-terminal IFT52 domain (residues 1-275; lane 4) alone, nor an IFT88/52 complex lacking the N-terminal IFT52 domain (with an IFT52(281-335) construct containing only the IFT88-binding region) (lane 5) show efficient pull-down of IFT-B2. Contaminants from the purification of the complex shown in lane 5 are marked with asterisks.

(B) Coomassie-stained SDS-PAGE gel showing a salt-stability test of the (IFT88/52N)-(IFT-B2) interaction. Pre-assembled IFT-B2 complex was bound to IFT88/52N-GST immobilized on GSH-beads. The beads were then divided into five aliquots which were washed with buffers containing increasing concentrations of NaCl. After these washes the bound material was eluted and analyzed. IFT172ΔC is the only salt-labile component in this complex and is already washed off at a NaCl-concentration of 150-200 mM, whereas all the other IFT-B2 proteins remain associated even after washing with 1 M NaCl.

(C) Coomassie-stained SDS-PAGE gels showing GST-pulldown experiments between IFT88/52N-GST complex immobilized on GSH beads and various combinations of IFT-B2 proteins. The left gel shows the IFT-B2 input mixtures, the right gel shows the bound material after washing and elution from the beads. A pentameric IFT-B2 complex (IFT80/57/54/38/20) is efficiently pulled down by IFT88/52N-GST, but not by empty beads (compare lanes 1 and 2). Omission of IFT54/20 (lane 4), or IFT80 (lane 5) does not influence the binding of any of the other components, but a lack of IFT57/38 in the mixture abolishes detectable pull-downs of IFT54/20 and IFT80 (lane 4), indicating that IFT57/38 is the direct interaction partner of IFT88/52N. Indeed, IFT57/38 alone (lane 7), but not IFT54/20 (lane 6) or IFT80 (lane 8), is sufficient to be pulled down in this assay. Contaminating bands from the IFT88/52N sample are marked with asterisks.
Intraflagellar transport proteins 172, 80, 57, 54, 38 and 20 form a stable tubulin-binding IFT-B2 complex (2016)
Figure 6

Figure 6. Crystal structure of the N-terminal domain of IFT52

(A) (top) Cartoon representation and surface plot of a superpositioning of CrIFT52N and MmIFT52N crystal structures. (bottom) Rotated 180 degrees compared to top panel. Conservation is mapped from 5 different species (see alignment in Appendix Fig. S5) and conserved residues are labeled (Cr numbering). The two residues (K130 and R204) found to disrupt interaction with IFT-B2 (panel (B)) are labeled in larger font.

(B) Coomassie stained SDS PAGE gel of a pull-down of IFT-B2 using WT or mutant IFT88/521-335-GST. Whereas single point mutations slightly reduced the amounts of pulled down IFT-B2, the K130E/R204E double mutation reduced it to background levels (compare lanes 2 and 6).
Histone methyltransferase DOT1L controls state-specific identity during B cell differentiation (2020)
Figure 1

Figure 1: Expression levels, efficient deletion of Dot1L in B cells and its effect on cellularity of B lineage subsets in the bone marrow.

A. Expression of Dot1L in different B-cell populations (Shi et al., 2015). B1: B1 cells, MZB: marginal zone B, FOB: follicular B, GCB: germinal center B, SPLPB: spleen plasma blast, SPLPC: spleen plasma cells, and BMPC: Bone marrow plasma cells. Expression is shown as transcript per million (TPM). Results represent the data from two biological replicates except SPLPB and BMPC which lack replicates (WT; n=2)..Bars indicate mean values.

B. Intracellular flow-cytometry staining for H3K79me2 in bone marrow B-cell subsets as well as splenic B and T cells from MB1Cre; Dot1Lwt/wt (WT) and MB1Cre; Dot1Lfl/fl (KO) mice. Results represent the data from two independent experiments.

C. Statistical analysis of absolute number of total nucleated cells from bone marrow B-cell subsets. Results represent the data pooled from three independent experiments and numbers represent biological replicates for each group (WT; n = 5, KO; n = 7). Bars and error bars indicate mean ± SD.

Data information: Statistical analyses were performed using the Student's two-tailed unpaired t-test. Statistical significance was determined by calculating the p-value. A p-value of less than 0.05 was considered as significant.