Journal: Skeletal Muscle

Article Title: SIX transcription factors are necessary for the activation of DUX4 expression in facioscapulohumeral muscular dystrophy

doi: 10.1186/s13395-024-00361-3

Figure Lengend Snippet: Low level of DUX4 expression in myoblasts are unaffected by SIX1/2/4 knockdown. FSHD1 (54–2) were transfected with SIX1, SIX2, and SIX4 siRNAs in proliferating myoblasts 72 h before harvesting. qPCR results show relative mRNA expression for A and C DUX4 , A-C DUX4 targets and D siRNAs targeting each respective SIX target. E-F Effects on SIX1 and SIX2 targets ( PGK1 , SLC4A7 and EYA1 ) were analyzed in E myoblasts and F myotubes. All conditions were carried out in triplicates with the mean and standard deviation depicted. Each experimental group ( n = 3) was normalized to a negative si-Control with the mean and standard deviation depicted. Asterisks demonstrate statistical significance between the control vs experimental group using an unpaired two tailed t- test (ns = not significant; * p < 0.05)

Article Snippet: SIX1 and SIX2 target genes and cofactors were used: PGK1 (Hs00943178_g1); SLC4A7 (Hs00186192_m1); EYA1 (Hs00166804_m1); EYA2 (Hs00193347_m1); EYA3 (Hs00544914_m1); EYA4 (Hs01012399_m1).

Techniques: Expressing, Knockdown, Transfection, Standard Deviation, Control, Two Tailed Test

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: Characterization of submandibular salivary glands in female and male Slc4a7 +/+ and Slc4a7 −/− mice

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Concentration Assay

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: Histology of submandibular glands (SMG) from wildtype and Slc4a7 null mice. Five μm thick sections of paraformaldehyde fixed mouse SMG were deparaffinized and stained with hematoxylin-eosin (HE): (a) female Slc4a7+/+ SMG, (b) female Slc4a7−/− SMG, (c) male Slc4a7+/+ SMG, and (d) male Slc4a7−/− SMG. Black scale bar = 100 μm. (e) The cross-sectional areas of acinar and ductal regions were calculated as described in Materials and Methods and expressed as the % acinar area. Acinar area (%) values are shown for individual glands (circles) along with the mean ± SEM. The cross-sectional areas of acinar and ductal regions were comparable in wildtype mice = 5 (2 males and 3 females) and Slc4a7 null mice = 6 (3 males and 3 females).

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Staining

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: Flow rate and amount of saliva in response to β-adrenergic vs. cholinergic stimulation of the ex vivo SMG. The mouse ex vivo SMG was stimulated for 20 min with 1.0 μM IPR, followed by 15 min washout, and finally 10 min perfusion with 0.3 μM CCh in physiological Solution A (Table 2). (a) No differences in the kinetics and rates of salivation (μl/min) were observed in response to 1.0 μM IPR or 0.3 μM CCh, respectively, in the SMG of Slc4a7+/+ (white circles, n = 11) and Slc4a7−/− (black circles, n = 15) mice. (b) The saliva amount (μl) of the ex vivo stimulated SMG in response to 1.0 μM IPR, and (c) in response to 0.3μM CCh were comparable in Slc4a7+/+ (white bars) and Slc4a7−/− mice (grey bars). Values are shown for individual glands (circles) along with the mean ± SEM.

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Ex Vivo

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: Flow rate and amount of saliva in response to physiological-like β-adrenergic and cholinergic stimulation of the ex vivo SMG. The mouse ex vivo SMG was stimulated for 10 min with the β-adrenergic receptor agonist IPR (1.0 μM) plus the cholinergic receptor agonist CCh (0.3 μM) in physiological Solution A (Table 2). (a) No differences in the kinetics and rates of salivation (μl/min) were observed between Slc4a7+/+ (white circle, n=14) and Slc4a7−/− mice (black circle, n=15). (b) The saliva amount (μl) of the ex vivo stimulated SMG was comparable in Slc4a7+/+ (white bars) and Slc4a7−/− mice (grey bars). Values are shown for individual glands (circles) along with the mean ± SEM.

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Ex Vivo

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: HCO3- concentration of saliva secreted by the ex vivo SMG. The HCO3- concentrations (mM) in the saliva of the ex vivo SMG were comparable in Slc4a7+/+ (white bars) and Slc4a7−/− mice (grey bars). As shown in Figures ​Figures33 and ​and4,4, salivation was stimulated with 1.0 μM IPR (circles: +/+, n=11; −/−, n=15; far left two bars), 0.3 μM CCh (diamonds: +/+, n=11; −/−, n=15; middle two bars), or 1.0 μM IPR plus 0.3 μM CCh (triangles: +/+, n=14; −/−, n=15; far right two bars). Values are shown for individual glands (symbols) along with the mean ± SEM.

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Concentration Assay, Ex Vivo

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: Mouse Slc4a7 expression in CHO-K1 cells mediates Na+-HCO3− cotransport. CHO-K1 cells transfected with either the full-length mouse Slc4a7 cDNA or empty vector were co-transfected with human CD8A cDNA and loaded with the pH sensitive dye BCECF in a K+ free solution (Solution B, Table 2) containing 0.1 mM of the Na+-K+ ATPase inhibitor Ouabain and Dynabeads™ CD8 beads. Extracellular Na+ was removed for at least 5 min, and then the intracellular alkalinization monitored following re-addition of extracellular Na+ in BCECF-loaded CHO-K1 cells decorated with CD8 beads. All solutions were Cl- free and contained 10 μM EIPA to block Cl-/HCO3- and Na+/H+ exchange, respectively. (a) A significant increase in the pHi was observed in Slc4a7 transfected CHO-K1 cells (black circles, n=10) when switched from Na+ free to Na+ containing (Solutions C and B, respectively, Table 2) HCO3- solutions. In contrast, an alkalinization was absent in empty vector transfected CHO-K1 cells (white circles, n=14). (b) Summary of the alkalinization rates for experiments like those shown in panel 6a for CHO-K1 cells transfected with either Slc4a7 (grey bars) or empty (white bars) vector. The Na+ dependent, Slc4a7 mediated alkalinization was completely inhibited either by the presence of 100 μM DIDS (4,4’-Diisothiocyano-2,2’-stilbenedisulfonic acid), 100 μM S-0859 (N-cyanosulphonamide) or by HCO3− free conditions in the presence of the carbonic anhydrase inhibitor ethoxzolamide (EZA, 30 μM), but was insensitive to 200 μM EIPA [results not shown, alkalinization rate (10−3/S): 200 μM EIPA = 2.55 ± 0.49 (n = 11) vs. 10 μM EIPA = 2.61 ± 0.41 (n = 10); p>0.92]. Values are shown for the average results (HCO3− containing = circles; DIDS = diamonds; S0859 = squares; HCO3− free = triangles) along with the mean ± SEM from at least ten different individual transfections. The rate of the Slc4a7 mediated alkalinization in a HCO3− containing solution (grey bar, black circles) was significantly greater (**, p<0.01) compared to all other conditions.

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Expressing, Transfection, Plasmid Preparation, Blocking Assay

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: Slc4a7 disruption fails to affect pHi regulation in SMG duct cells. Duct cells isolated from Slc4a7+/+ and Slc4a7−/− mouse SMG were loaded with the pH sensitive dye BCECF and the Na+ dependent intracellular pH (pHi) recovery monitored following an NH4+-prepulse induced intracellular acidosis. All solutions were Cl- free and contained 10 μM EIPA to block Cl-/HCO3- and Na+/H+ exchange, respectively. HCO3- containing and HCO3- free NH4+ solutions were Solutions E and H (Table 2), respectively. (a) The increase in the pHi observed in duct cells of Slc4a7−/− (black circles, n=15) and Slc4a7+/+ (white circles, n=13) mice was comparable when switched from Na+ free to Na+ containing HCO3- solutions (Solutions F and D, respectively, Table 2). (b) Summary of the alkalinization rates for experiments like those shown in panel 7a for SMG duct cells from either Slc4a7−/− (grey bars) or Slc4a7+/+ (white bars) mice. Values are shown for individual mice (HCO3− containing = circles; DIDS = diamonds; S0859 = squares; HCO3− free = triangles) along with the mean ± SEM of ducts isolated from at least six different mice. The alkalinization rate in a HCO3− containing solution for Slc4a7+/+ mice (white bar, white circles) was significantly greater (*, p<0.05) than in the presence of DIDS (white bar, white diamonds). Of note, the Na+ dependent alkalinization induced in SMG duct cells by switching from Solution F to D (Table 2) was only moderately sensitive to DIDS (100 μM) and S0859 (100 μM) or dependent on HCO3- (switching from Na+ free to Na+ containing HCO3- free solutions, Solution I to G, Table 2, HCO3− free in the presence of 30 μM carbonic anhydrase inhibitor EZA) [results not shown for 500 μM DIDS, alkalinization rate (10−3/S): 500 μM DIDS = 1.83 ± 0.40 (n = 7) and 2.12 ± 0.27 (n = 10) for Slc4a7+/+ and Slc4a7−/−, respectively; ANOVA test p>0.66]. (c) Summary of qPCR results for Slc4 (a2, a4, a5, a8, a9, a10 and a11), Slc9 (a1, a2 and a3) and Cftr, normalized to β-actin expression. Values are shown for individual SMG (white circles = Slc4a7+/+; black circles = Slc4a7−/−) along with the mean ± SEM of glands isolated from 4 Slc4a7+/+ and 6 Slc4a7−/− mice. Note break in Y axis for Slc9a1 and Cftr.

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Isolation, Blocking Assay, Expressing

Journal: Journal of cellular physiology

Article Title: The Apical Na + -HCO 3 - Cotransporter Slc4a7 (NBCn1) does not contribute to Bicarbonate Transport by Mouse Salivary Gland Ducts

doi: 10.1002/jcp.28306

Figure Lengend Snippet: Immunolocalization of Slc4a7 in the mouse SMG. Immunofluorescence staining of Slc4a7 (green) and Slc12a2/Nkcc1 (red), while nuclei were stained with DAPI (blue). (a) Slc4a7 localized at the apical membrane (white arrowhead) with faint, diffuse intracellular staining in the SMG ducts of Slc4a7+/+ mice, but no immunostaining was detected in acinar cells. (b) Slc12a2 was labelled at the basolateral membranes of acinar cells from Slc4a7+/+ mice (yellow arrow). (c) Merged image of Slc4a7 (white arrowhead) and Slc12a2 (yellow arrow) staining in Slc4a7+/+ mice. (d) Slc4a7-specific immunostaining was not detected in the SMG ducts of the Slc4a7−/− mice. (e) Slc12a2 immunostaining was unchanged in Slc4a7−/− mice (yellow arrow). (f) Merged image of Slc4a7 and Slc12a2 (yellow arrow) staining from Slc4a7−/− mice. White scale bar = 10 μm; immunostaining patterns were comparable in wildtype mice = 5 (2 males and 3 females) and Slc4a7 null mice = 6 (3 males and 3 females). Images shown are from female Slc4a7+/+ and Slc4a7−/− mice.

Article Snippet: Sections were blocked in 1% BSA in PBS (phosphate buffered saline) for 30 min at RT, and then incubated at 4°C with rabbit anti-Slc4a7 (1:100 dilution, OriGene, Cat# TA322834, RRID: AB_2749851) and goat anti-Slc12a2 (NKCCl, 1:100 dilution, Santa Cruz Biotechnology, Cat# sc-21545, RRID: AB_2188633) antibodies overnight.

Techniques: Immunofluorescence, Staining, Immunostaining

Journal: Cell Host & Microbe

Article Title: The Bicarbonate Transporter SLC4A7 Plays a Key Role in Macrophage Phagosome Acidification

doi: 10.1016/j.chom.2018.04.013

Figure Lengend Snippet: SLC-Focused CRISPR/Cas9 Genetic Loss-of-Function Screens Identify SLC4A7 to Be Important for Phagosome Acidification (A) Schematic representation of the major steps of the SLC-focused CRISPR/Cas9 screen to identify SLCs involved in phagocytosis. (B) Representative flow cytometry scatterplot of phagocytosis assays. PMA-differentiated U937 cells were incubated with dual-colored opsonized beads. Each dot represents one cell: intensity of the pH-insensitive dye (YG) is displayed on the x axis, intensity of pH-sensitive dye (pHrodo-Red), whose signal intensity increases with decrease in pH, is shown on the y axis. Double-negative cells were classified as phagocytosis-negative (PhagoNeg), double-positive cells (YG and high pHrodo-Red signal) were classified as cells having undergone phagocytosis and phagosome acidification (PhagoLate), and single positive cells (YG and low pHrodo-Red signal) were classified as cells at early stages of phagocytosis (PhagoEarly). The marginal intensity distributions are shown on the sides of the plot. (C) Volcano plot showing the statistical significance of genes depleted in the PhagoLate population on the y axis as –log 10 (p adj ) against average log 2 fold-change (mean LFC) on the x axis calculated for all sgRNAs per gene. The size of the dots represents the number of significant changes in sgRNAs counts. See Figure S1 B for differential abundance of individual sgRNAs. (D) Immunoblot analysis of primary human monocytes (Mo) derived from peripheral blood and M-CSF-differentiated monocyte-derived human macrophages, which were unpolarized (M0), or polarized toward M1 phenotype with interferon-γ and lipopolysaccharide, or to M2 phenotype with interleukin-4. Respective lysates were probed with an anti-SLC4A7 antibody detecting all isoforms (C-terminal epitope) and an anti-SLC4A7 antibody detecting an N-terminal epitope present only in isoforms starting with the amino acids “MEAD.” Actin was used as loading control. MW, molecular weight. See Figure S1 C for isoforms. (E) Representative immunoblot analysis of SLC4A7 expression in undifferentiated and PMA-differentiated THP-1 and U937 cells. Actin was used as loading control. MW, molecular weight. (F) Representative kinetics of phagosome acidification in four independent SLC4A7 knockout (sg1-sg4) and control (sgRen) U937 cells. After PMA differentiation, U937 cells were incubated with dual-colored beads and analyzed at the indicated time points using flow cytometry. The fraction of PhagoLate cells is displayed against the incubation time with beads. See Figure S1 D for confirmation of SLC4A7 knockdown. (G) Phagocytosis assays with PMA-differentiated SLC4A7 knockout (sg1) or control (sgRen) THP-1 cells, which were incubated with dual-colored beads as described in ). Bar graphs show the fraction of PhagoLate, PhagoEarly, and PhagoNeg cells as assessed by pHrodo and YG fluorescence in flow cytometry. Data are mean ± 95% confidence interval from four replicates. ∗ p < 0.05, ∗∗ p < 0.001; by Welsh's t test.

Article Snippet: Codon optimized SLC4A7 isoform 1 and 6 cDNA , GenScript , N/A.

Techniques: CRISPR, Flow Cytometry, Incubation, Western Blot, Derivative Assay, Control, Molecular Weight, Expressing, Knock-Out, Knockdown, Fluorescence

Journal: Cell Host & Microbe

Article Title: The Bicarbonate Transporter SLC4A7 Plays a Key Role in Macrophage Phagosome Acidification

doi: 10.1016/j.chom.2018.04.013

Figure Lengend Snippet: Functional Consequences of SLC4A7 Knockout and Overexpression (A) Phagocytosis assays with control (sgRen) and SLC4A7 knockout (sg1, sg4) U937 cells, which were lentivirally infected to exogenously express HA-tagged SLC4A7 isoform 1, isoform 6, or empty vector control (Ctrl), respectively. Cells were incubated with dual-colored beads as described in Figure 1 B. Bar graphs show the fraction of PhagoLate, PhagoEarly, and PhagoNeg cells as assessed by flow cytometry. Data are mean ± 95% confidence interval from three replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; by Welsh's t test. See Figure S1 F for immunoblot confirmation of SLC4A7 knockout and exogenous expression. (B) Phagocytosis assays with control (sgRen) and SLC4A7 knockout (sg1) THP-1 cells, which were lentivirally infected to exogenously express HA-tagged SLC4A7 isoform 1, isoform 6, or empty vector control (Ctrl), respectively. Cells were incubated with dual-colored beads as in B and A. Bar graphs show the fraction of PhagoLate, PhagoEarly, and PhagoNeg cells as assessed by flow cytometry. Data are mean ± SD from two replicates. ∗ p < 0.05, ∗∗ p < 0.01; by Welsh's t test. See Figure S1 G for immunoblot confirmation of SLC4A7 knockdown and exogenous expression. (C) Intracellular killing assay with viable Gram-negative ( E. coli ) and Gram-positive ( Streptococcus pyogenes ΔSLO, Staphylococcus carnosus Schleifer and Fischer, and Staphylococcus aureus Newman and USA300) bacteria in control (sgRen), SLC4A7 knockout (sg1), and SLC4A7 knockout reconstituted with SLC4A7 isoform 6 (sg1-SLC4A7(i6)) THP-1 cells. Bar graphs depict the percentage of surviving intracellular bacteria in relation to time point zero. Data are median and interquartile range from three replicates. ns, not significant, ∗∗∗ p < 0.001; by Wilcoxon-Mann-Whitney test. (D) Representative confocal immunofluorescence images of endogenous SLC4A7 in control (sgRen) or SLC4A7 knockout (sg1) THP-1 cells. PMA-differentiated cells were fixed and stained with anti-SLC4A7 antibody (green). DNA was counterstained with DAPI (blue). The overlay of both signals is depicted. Scale bars, 5 μm. (E) Representative confocal live-cell immunofluorescence images of THP-1 cells expressing GFP-tagged SLC4A7 isoform 6. After PMA-induced differentiation, cells were incubated with pHrodo-labeled heat-killed S. aureus (HKSA, upper panel) or dual-colored beads (pHrodo and bright blue; lower panel). Single channel images and respective overlays are shown. Scale bars, 10 μm. For time-lapse acquisitions, see . (F) Simultaneous measurement of cytoplasmic and phagosomal pH during phagocytosis using live-cell microscopy. PMA-differentiated control (sgRen) and SLC4A7 knockout (sg1) THP-1 cells were loaded with BCECF-AM, incubated with dual-colored beads (pHrodo and bright blue), and imaged at the indicated time points. Incubation and imaging were done in Hank’s balanced salt solution with 10% FCS at 37°C in 5% CO 2 . At each time point, z stacks of five different fields were acquired per replicate. Bar charts represent pHrodo intensities of phagocytosed beads or cytoplasmic pH as calculated based on the BCECF calibration curve. Data are mean and 95% confidence interval from three replicates. ∗∗∗ p < 0.001; by Welch's t test. For in situ calibration of the BCECF 490/440 ratio, see Figure S2 A; for example images, see Figure S2 B. For simultaneous cytoplasmic and phagosomal pH measurements in THP-1 cells phagocyting heat-killed S. aureus , see Figure S2 C (left panel), and for U937 cells phagocytosing beads, see Figure S2 C (right panel). (G) Schematic representation of the SLC4A7 model with the transmembrane domains (TMDs) of the core domain in yellow and the TMDs of the gate domain in blue. Helix 3 and helix 10, which align to form a continuous helix near the putative substrate binding site, are shown in orange and green, respectively. The residues mutated in the functional studies are shown in red with the isoform1/isoform6 numbering scheme. (H) Phagocytosis assays with two independent U937 clones with SLC4A7 knockout (sg1, sg4) or control (sgRen), which were infected with lentiviral expression constructs coding for Strep-HA-tagged SLC4A7 isoform 6 (SLC4A7(i6)), an isoform 6 mutant lacking amino acids 1,008–1,131 (delC), two different predicted transport mutants (T549I and D811A), or empty vector control (Ctrl). PMA-differentiated cells were incubated with dual-colored beads as in Figure 1 B and analyzed by flow cytometry. Bar graphs show the fraction of PhagoLate cells as assessed by pHrodo fluorescence intensity. Data are mean and 95% confidence interval from two replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; by Welsh's t test. For subcellular localization of the different proteins, see immunofluorescence analysis in Figure S2 E.

Article Snippet: Codon optimized SLC4A7 isoform 1 and 6 cDNA , GenScript , N/A.

Techniques: Functional Assay, Knock-Out, Over Expression, Control, Infection, Plasmid Preparation, Incubation, Flow Cytometry, Western Blot, Expressing, Knockdown, Bacteria, MANN-WHITNEY, Immunofluorescence, Staining, Labeling, Microscopy, Imaging, In Situ, Binding Assay, Clone Assay, Construct, Mutagenesis, Fluorescence

Journal: Cell Host & Microbe

Article Title: The Bicarbonate Transporter SLC4A7 Plays a Key Role in Macrophage Phagosome Acidification

doi: 10.1016/j.chom.2018.04.013

Figure Lengend Snippet:

Article Snippet: Codon optimized SLC4A7 isoform 1 and 6 cDNA , GenScript , N/A.

Techniques: Virus, Recombinant, Software, Protease Inhibitor, Western Blot, Mutagenesis