rabbit anti αβ crystallin  (Cell Signaling Technology Inc)

Journal: BMC Biology

Article Title: Guanine quadruplexes mediate mitochondrial RNA polymerase pausing

doi: 10.1186/s12915-025-02229-4

Figure Lengend Snippet: POLRMT pausing impairs renal proximal tubule function. A Confocal image showing RHPS4 (green) uptake into mitochondria following 4 and 24 h of drug treatment. Mitochondria are stained with Mitotracker (grey). Cells in the upper panels are not stained with MitoTracker. Scale bar is 10 µm. B Mitochondrial gene expression in renal proximal tubule epithelial cells (RPTEC) 24 h after RHPS4 treatment, normalized to vehicle treated cells. Gene expression is in arbitrary units ( n = 3; P < 0.0001, one-way ANOVA, error bars = S.E.M.). C mtDNA copies per nuclear genome ( n = 3; P > 0.05, t-test) ( D ) mtDNA copies per total DNA mass ( n = 3; P > 0.05, t-test). E Oxygen consumption (OCR) is lower ( n = 5; P < 0.0001, two-way ANOVA, error bars = S.D.) and ( F ) Extracellular acidification rate (ECAR) is higher ( n = 5, P < 0.0001, two-way ANOVA, error bars = S.D.) in RPTEC 24 h after treatment with RHPS4. G Protein expression of nuclear-encoded TOMM20, mitochondria-encoded COX1 ( MT-CO1 ) and AMPK phosphorylation before and after 72 h of RHPS4 treatment. H Schematic of renal proximal tubule culture system, where cells are seeded on the bottom of the permeable membrane. I Active transport of glucose analogue 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)−2-Deoxyglucose (2-NBDG), expressed as a percentage of transport performed by DMSO-treated cells ( n ≥ 5; *** P < 0.001, t-test, error bars = S.E.M)

Article Snippet: Primary antibodies were diluted 1:1000 in blocking buffer and included rabbit α-TOMM20 (Cell Signaling Technologies), mouse α-COX1 (AbCam), rabbit α-phospho-AMPK (Thr172) (Cell Signaling Technologies), SGLT2 (Abcam), and rabbit α-GAPDH (AbCam) as a loading control.

Techniques: Staining, Gene Expression, Expressing, Phospho-proteomics, Membrane

Journal: eLife

Article Title: Flamingo participates in multiple models of cell competition

doi: 10.7554/eLife.98535

Figure Lengend Snippet: ( A–C ) Ras V12 , scrib RNAi tumors stained for DAPI, Dcp1+ ( A ), and puc-lacZ ( B ). ( C ) Merged channels. ( D ) Representation of how Dcp1+ staining localizes in the wildtype (WT) cells at the boundary with the tumor. ( E–G ) Ras V12 , scrib RNAi tumors mutant for Fmi, stained with DAPI, Dcp1+ ( E ), and puc-LacZ ( F ). ( G ) Merged channels. ( H ) Representation of how Dcp1+ staining localizes in the tumor cells in contact with the surrounding WT tissue. Scale bar for A–G: 25 μm. ( I–K ) Dcp1+ staining in >>Myc clones in eye discs. >>Myc clones are marked by GFP ( I ) and were stained against Dcp1+ ( J ). The arrows show apoptotic WT (red arrow) and >>Myc (white arrow) cells at the clone boundary. ( K ) Merged channels, showing apoptotic cells evenly distributed between WT and >>Myc cells. ( L–N ) Dcp1+ staining in >>Myc clones lacking Fmi in the eye disc. Eye disc >>Myc clones are marked by GFP ( L ) and were stained for Dcp1+ ( M , N ) Merged channels, showing apoptotic cells localized mainly in the >>Myc, fmi E59 clones. Scale bar: 50 μm. ( O ) Quantification of apoptotic cells in WT vs >>Myc clones in eye discs. Apoptosis occurs similarly in WT and >>Myc cells (two-tailed paired t-test; p-value = 0.6049). The left side of the graph shows the number of apoptotic WT and >>Myc cells. Each imaginal disc is displayed as a pair of dots, linked by a line, to easily visualize the Dcp1+ apoptotic cells in WT vs >Myc cells. Dots represent the number of apoptotic WT (left) or >>Myc (right) cells per disc. The right side of the graph displays the difference (>>Myc minus WT apoptotic cells). The dashed line indicates the mean difference between those values for all samples. N=14 discs. ( P ) Quantification of apoptotic cells in WT vs >>Myc, fmi E59 clones in eye discs. Apoptosis is found mainly in >>Myc, fmi E59 cells (two-tailed paired t-test; p-value = 0.0006). The left side of the graph shows the number of apoptotic WT and >>Myc, fmi E59 cells, side by side. Dots represent the number of apoptotic >>WT (left) or >>Myc, fmi E59 (right) cells per disc. The right side of the graph displays the difference (>>Myc, fmi E59 minus WT apoptotic cells). The dashed line indicates the mean difference between those values for all samples. N=14 discs. ( Q–R ) Proliferation analysis performed by pHis3 staining in wing discs with either >>Myc clones ( Q ) or >>Myc, fmi E59 clones ( R ). Scale bar: 20 μm. ( S ) Proliferative ratio of GFP cells in a non-competition Control (n=9 discs), >>Myc (n=9 discs), or >>Myc, fmi E59 (n=13 discs) clones. The proliferative ratio for each group was calculated as the ratio of pHis3 cells within the GFP+ clone vs the non-GFP WT tissue and the differences were analyzed as an ordinary ANOVA with a Tukey’s test for multiple comparisons, with all p-values<0.0001 (****).

Article Snippet: We used the following primary antibodies: rabbit α-pHis3 (Millipore), 1:100; rabbit α-Dcp1 (Cell Signaling), 1:100.

Techniques: Staining, Mutagenesis, Clone Assay, Two Tailed Test, Control